PCB153-Induced Overexpression of ID3 Contributes to the Development of Microvascular Lesions
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{"title"=>"PCB153-induced overexpression of ID3 contributes to the development of microvascular lesions", "type"=>"journal", "authors"=>[{"first_name"=>"Jayanta K.", "last_name"=>"Das", "scopus_author_id"=>"14033707900"}, {"first_name"=>"Quentin", "last_name"=>"Felty", "scopus_author_id"=>"8527096000"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"373700480", "sgr"=>"84905474540", "issn"=>"19326203", "pmid"=>"25090023", "scopus"=>"2-s2.0-84905474540", "doi"=>"10.1371/journal.pone.0104159"}, "id"=>"ca4189bd-14bf-3628-907d-5a24565ed386", "abstract"=>"Microvascular lesions resulting from endothelial cell dysfunction are produced in the brain, lung, kidney, and retina of patients of complex chronic diseases. The environmental and molecular risk factors which may contribute in the development of microvascular damage are unclear. The mechanism(s) responsible for initiating microvascular damage remain poorly defined, although several inciting factors have been proposed, including environmental toxicants-induced oxidative stress. Enhanced neovascularization has been implicated in either the development or progression of proliferative vascular lesions. Here, we present evidence for how PCB-induced ROS may contribute to the development of a neovascular phenotype with the aim of elucidating the role of environmental toxicants in endothelial dysfunction with a specific focus on the inhibitor of differentiation protein ID3. We used a combination of phenotype and immunohistochemical analysis followed by validating with protein expression and post-translational modifications with Western Blot and MALDI-TOF/TOF analysis. We also looked for a correlation between ID3 expression in vascular tissue. Our results showed that PCB-induced ROS mediated a highly tube branched neovascular phenotype that also depended on ID3 and Pyk2; and PCB153 treatment increased the size of endothelial spheroids under conditions typically used for clonal selection of stem cell spheroids. High ID3 protein expression correlated with a greater degree of malignancy and oxidative DNA damage marker 8-OHdG in blood vessels from human subjects. PCB153 treatment increased both serine and tyrosine phosphorylation of endothelial ID3. Stable ID3 overexpression increased cell survival of human microvascular endothelial cell line hCMEC/D3. In summary, our data provide evidence that ID3 may play a critical role in regulating vascular endothelial cell survival and development of microvascular lesions induced by persistent environmental pollutants such as PCB153. Findings of this study are important because they provide a new paradigm by which PCBs may contribute to the growth of microvascular lesions.", "link"=>"http://www.mendeley.com/research/pcb153induced-overexpression-id3-contributes-development-microvascular-lesions", "reader_count"=>8, "reader_count_by_academic_status"=>{"Student > Ph. D. Student"=>4, "Student > Master"=>3, "Student > Bachelor"=>1}, "reader_count_by_user_role"=>{"Student > Ph. D. Student"=>4, "Student > Master"=>3, "Student > Bachelor"=>1}, "reader_count_by_subject_area"=>{"Environmental Science"=>2, "Agricultural and Biological Sciences"=>2, "Medicine and Dentistry"=>1, "Neuroscience"=>1, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Chemistry"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Neuroscience"=>{"Neuroscience"=>1}, "Chemistry"=>{"Chemistry"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>2}, "Environmental Science"=>{"Environmental Science"=>2}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "reader_count_by_country"=>{"United States"=>1}, "group_count"=>1}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1620955"], "description"=>"<p>MALDI-TOF/TOF spectra for peptides extracted from hCMEC/D3 cells. Vehicle control showed that Ser-5 or Ser-18 (<i>Panel </i><b><i>A</i></b>) and Tyr-11 (<i>Panel </i><b><i>B</i></b>) are phosphorylated. Identification of phosphopeptides from PCB153 treated cells indicated that along with Ser-5 that was detected at the level of MS, Tyr-48 was also phosphorylated (<i>Panel </i><b><i>C</i></b>). Methods: Cell lysate was immunoprecipitated with ID3 antibody from vehicle control and PCB153 treated hCMEC/D3 cells. The immunoprecipitated lysate was separated by 1D-SDS-PAGE and the protein band corresponding to the MW of ID3 was excised from the gel, subjected to tryptic digestion and phosphopeptide enrichment. The phosphopeptides were spotted on a MALDI plate followed by MS and database search. The spectra of all peptides were manually evaluated for the loss of phosphate which is shown in red circles and the observed mass (dashed circle) and sequence of the peptides are shown. <i>X-axis</i> represents mass and <i>Y-axis</i> represents intensity.</p>", "links"=>[], "tags"=>["cell biology", "Cell processes", "Cell proliferation", "oxidative stress", "physiology", "Cardiovascular physiology", "angiogenesis", "Public and occupational health", "Environmental health", "Vascular medicine", "Vascular diseases", "phosphorylated", "amino", "acids", "id3", "microvascular"], "article_id"=>1126912, "categories"=>["Biological Sciences"], "users"=>["Jayanta K. Das", "Quentin Felty"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0104159.g006", "stats"=>{"downloads"=>4, "page_views"=>22, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Identification_of_phosphorylated_amino_acids_in_ID3_from_microvascular_ECs_/1126912", "title"=>"Identification of phosphorylated amino acids in ID3 from microvascular ECs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-08-04 03:13:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/1620958"], "description"=>"<p>hCMEC/D3 cells were treated with PCB153 to determine its effect on the tyrosine phosphorylation state of ID3 at 6, 12, and 24 h. After 24 h of PCB153 [10 µM] treatment, we observed a significantly higher level of tyrosine phosphorylation than vehicle control. The phospho-Tyr bands detected from immunoprecipitated (IP) cell lysate corresponded with the molecular weight (MW) of 17 kDa for ID3. IP was performed to isolate ID3 from cell lysate using a monoclonal anti-ID3 antibody. Magnetic bead-based separation was then used to extract the bound ID3 protein via Protein G coupled to superparamagnetic Dynabeads. P-Tyr was detected by immunoblotting using the anti-phospho-Tyr antibody. Electrochemiluminescence (ECL) band intensity for phospho-Tyr was imaged with a Bio-Rad Versa Doc instrument.</p>", "links"=>[], "tags"=>["cell biology", "Cell processes", "Cell proliferation", "oxidative stress", "physiology", "Cardiovascular physiology", "angiogenesis", "Public and occupational health", "Environmental health", "Vascular medicine", "Vascular diseases", "tyrosine", "phosphorylation"], "article_id"=>1126915, "categories"=>["Biological Sciences"], "users"=>["Jayanta K. Das", "Quentin Felty"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0104159.g007", "stats"=>{"downloads"=>3, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_PCB153_increased_tyrosine_phosphorylation_of_ID3_/1126915", "title"=>"PCB153 increased tyrosine phosphorylation of ID3.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-08-04 03:13:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/1620960"], "description"=>"<p>PCB-induced vessel formation was characterized in a co-culture model in which HUVECs were seeded on top of a confluent layer of fibroblasts (Fb). HUVECs were transiently transfected with either Pyk2 siRNA [50 nM] or Silencer negative control siRNA for 48 h prior to experiments. Co-cultures immunostained for endothelial marker CD31 showed a robust inhibition of PCB-induced neovascular network by Pyk2 siRNA when compared to cells transfected with negative control siRNA. Scrambled siRNA (Silencer negative control siRNA, Ambion) was used in all RNAi experiments. The data shown are representative images from an experiment repeated three separate times. HUVECs were treated with 1 ng/ml of PCB153 (2,2′,4,4′,5,5′-hexachlorobiphenyl) or PCB77 (3,3′,4,4′-tetrachlorobiphenyl).</p>", "links"=>[], "tags"=>["cell biology", "Cell processes", "Cell proliferation", "oxidative stress", "physiology", "Cardiovascular physiology", "angiogenesis", "Public and occupational health", "Environmental health", "Vascular medicine", "Vascular diseases", "pyk2", "inhibited", "pcb-induced", "neovascular"], "article_id"=>1126917, "categories"=>["Biological Sciences"], "users"=>["Jayanta K. Das", "Quentin Felty"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0104159.g008", "stats"=>{"downloads"=>1, "page_views"=>17, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_siRNA_Pyk2_inhibited_the_PCB_induced_neovascular_phenotype_/1126917", "title"=>"siRNA Pyk2 inhibited the PCB-induced neovascular phenotype.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-08-04 03:13:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/1620961"], "description"=>"<p>(<b>A</b>) hCMEC/D3 cells were treated with PCB153 to determine its effect on Pyk2 phosphorylation and whether the effect depended on ROS. PCB153-induced ROS increased the level of pY<sup>402</sup> Pyk2 which was inhibited by the ROS scavenger N-acetylcysteine. (<b>B</b>) To determine whether ID3 was a downstream target of functional Pyk2 kinase, ECs were treated with the chemical Pyk2 kinase inhibitor PF-431396 [1 µM]. Pyk2 inhibitor inhibited the level of pY<sup>402</sup> Pyk2, but did not change ID3 expression compared to vehicle control. (<b>C</b>) Using a loss-of-function approach with siRNA Pyk2, we compared the effects of inhibiting Pyk2 on the total level of ID3 protein. Using Lipofectamine RNAiMAX reagent (Invitrogen) ECs were transfected with either Pyk2 siRNA or Silencer negative control siRNA for 48 h prior to PCB153 treatments. Pyk2 siRNA inhibited total ID3 protein levels in both the PCB153 and vehicle control treated hCMEC/D3 cells compared to the negative control. Pyk2 siRNA treatment also inhibited the level of phosphotyrosine that corresponded to MW∼17 kDa of ID3.</p>", "links"=>[], "tags"=>["cell biology", "Cell processes", "Cell proliferation", "oxidative stress", "physiology", "Cardiovascular physiology", "angiogenesis", "Public and occupational health", "Environmental health", "Vascular medicine", "Vascular diseases", "pyk2", "phosphorylation", "regulated", "id3"], "article_id"=>1126918, "categories"=>["Biological Sciences"], "users"=>["Jayanta K. Das", "Quentin Felty"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0104159.g009", "stats"=>{"downloads"=>5, "page_views"=>43, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_PCB153_increased_Pyk2_phosphorylation_amp_Pyk2_regulated_ID3_protein_/1126918", "title"=>"PCB153 increased Pyk2 phosphorylation & Pyk2 regulated ID3 protein.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-08-04 03:13:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/1620962"], "description"=>"<p>Pyk2 siRNA significantly inhibited tyrosine phosphorylation for proteins of MW∼17 kDa corresponding to ID3 therefore we determined whether ID3 was phosphorylated by recombinant active Pyk2 in an <i>in vitro</i> kinase assay. Pyk2 kinase was incubated with either Flag-tagged ID3 or GST-tagged ID3 in the presence or absence of ATP; and phosphorylation was detected by immunoblotting. (<b>A</b>) A phosphorylated tyrosine band was detected at the corresponding MW∼13 kDA for FLAG-ID3 and MW∼39 kDa for GST-ID3. (<b>B</b>) Phosphorylation of in vitro–phosphorylated FLAG-ID3 was confirmed by combined MS+MS/MS as described previously. MS spectra analysis identified the human ID3 sequence with the protein score of 100%. Presence of phosphorylated amino acids was identified by the MS/MS peak showing neutral loss of phosphates which is shown in red circles and the observed mass (dashed circle). The sequence of the peptides are shown. <i>X-axis</i> represents mass and <i>Y-axis</i> represents intensity.</p>", "links"=>[], "tags"=>["cell biology", "Cell processes", "Cell proliferation", "oxidative stress", "physiology", "Cardiovascular physiology", "angiogenesis", "Public and occupational health", "Environmental health", "Vascular medicine", "Vascular diseases", "pyk2", "kinase", "phosphorylated"], "article_id"=>1126919, "categories"=>["Biological Sciences"], "users"=>["Jayanta K. Das", "Quentin Felty"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0104159.g010", "stats"=>{"downloads"=>1, "page_views"=>15, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Recombinant_active_Pyk2_kinase_phosphorylated_ID3_/1126919", "title"=>"Recombinant active Pyk2 kinase phosphorylated ID3.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-08-04 03:13:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/1620963"], "description"=>"<p>The functional significance of ID3 on survival of hCMEC/D3 cells was determined by generating a stable ID3 overexpressing cell line. (<b>A</b>) and (<b>B</b>) Confirmation of ID3 overexpression in a stable clone of hCMEC/D3 cells was shown by both immunoflouorscence microscopy and immunoblot of ID3 levels. (<b>C</b>) Cells were subjected to FACS analysis after FITC Annexin V (<i>Y-axis</i>) and Propidium Iodide (PI: <i>X-axis</i>) staining. ID3 clone indicated a greater percentage of live cells stained negative for both FITC Annexin V and PI. ID3 clone showed 88.74% of cell population was not apoptotic compared to a far lower percentage of 25.74% live cells in the control.</p>", "links"=>[], "tags"=>["cell biology", "Cell processes", "Cell proliferation", "oxidative stress", "physiology", "Cardiovascular physiology", "angiogenesis", "Public and occupational health", "Environmental health", "Vascular medicine", "Vascular diseases", "overexpression"], "article_id"=>1126920, "categories"=>["Biological Sciences"], "users"=>["Jayanta K. Das", "Quentin Felty"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0104159.g011", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_ID3_overexpression_increased_cell_survival_of_ECs_/1126920", "title"=>"ID3 overexpression increased cell survival of ECs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-08-04 03:13:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/1620964"], "description"=>"<div><p>Microvascular lesions resulting from endothelial cell dysfunction are produced in the brain, lung, kidney, and retina of patients of complex chronic diseases. The environmental and molecular risk factors which may contribute in the development of microvascular damage are unclear. The mechanism(s) responsible for initiating microvascular damage remain poorly defined, although several inciting factors have been proposed, including environmental toxicants-induced oxidative stress. Enhanced neovascularization has been implicated in either the development or progression of proliferative vascular lesions. Here, we present evidence for how PCB-induced ROS may contribute to the development of a neovascular phenotype with the aim of elucidating the role of environmental toxicants in endothelial dysfunction with a specific focus on the inhibitor of differentiation protein ID3. We used a combination of phenotype and immunohistochemical analysis followed by validating with protein expression and post-translational modifications with Western Blot and MALDI-TOF/TOF analysis. We also looked for a correlation between ID3 expression in vascular tissue. Our results showed that PCB-induced ROS mediated a highly tube branched neovascular phenotype that also depended on ID3 and Pyk2; and PCB153 treatment increased the size of endothelial spheroids under conditions typically used for clonal selection of stem cell spheroids. High ID3 protein expression correlated with a greater degree of malignancy and oxidative DNA damage marker 8-OHdG in blood vessels from human subjects. PCB153 treatment increased both serine and tyrosine phosphorylation of endothelial ID3. Stable ID3 overexpression increased cell survival of human microvascular endothelial cell line hCMEC/D3. In summary, our data provide evidence that ID3 may play a critical role in regulating vascular endothelial cell survival and development of microvascular lesions induced by persistent environmental pollutants such as PCB153. Findings of this study are important because they provide a new paradigm by which PCBs may contribute to the growth of microvascular lesions.</p></div>", "links"=>[], "tags"=>["cell biology", "Cell processes", "Cell proliferation", "oxidative stress", "physiology", "Cardiovascular physiology", "angiogenesis", "Public and occupational health", "Environmental health", "Vascular medicine", "Vascular diseases", "overexpression", "id3", "contributes", "microvascular"], "article_id"=>1126921, "categories"=>["Biological Sciences"], "users"=>["Jayanta K. Das", "Quentin Felty"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0104159", "stats"=>{"downloads"=>10, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_PCB153_Induced_Overexpression_of_ID3_Contributes_to_the_Development_of_Microvascular_Lesions_/1126921", "title"=>"PCB153-Induced Overexpression of ID3 Contributes to the Development of Microvascular Lesions", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2014-08-04 03:13:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/1620940"], "description"=>"<p>Representative micrographs showing inhibition of endothelial tube formation in HUVECs cultured in a matrix gel and exposed for 5(<i>Panel </i><b><i>A</i></b>). Co-treatments with antioxidant enzymes MnSOD (<i>Panel </i><b><i>B</i></b>) and catalase (<i>Panel </i><b><i>C</i></b>) were used to determine the contribution of specific ROS. HUVECs were treated with vehicle control (0.1% DMSO) or 1 ng/ml of PCB153 and PCB77 for 5 h. For antioxidant enzyme overexpression, prior to treatment HUVECs were incubated with replication deficient adenoviral constructs containing human MnSOD, catalase, or empty vector (ViraQuest Inc.) at 50 multiplicities of infection (MOI) for 24 h. HUVECs were treated with 1 ng/ml of PCB153 (2,2′,4,4′,5,5′-hexachlorobiphenyl) or PCB77 (3,3′,4,4′-tetrachlorobiphenyl).</p>", "links"=>[], "tags"=>["cell biology", "Cell processes", "Cell proliferation", "oxidative stress", "physiology", "Cardiovascular physiology", "angiogenesis", "Public and occupational health", "Environmental health", "Vascular medicine", "Vascular diseases", "neovascularization", "phenotype", "depends"], "article_id"=>1126897, "categories"=>["Biological Sciences"], "users"=>["Jayanta K. Das", "Quentin Felty"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0104159.g001", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_PCB_induced_neovascularization_phenotype_depends_on_ROS_/1126897", "title"=>"PCB-induced neovascularization phenotype depends on ROS.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-08-04 03:13:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/1620941"], "description"=>"<p>PCB-induced vessel formation was further characterized in a co-culture model. HUVECs were seeded on top of a confluent layer of fibroblasts (Fb). (<b>A</b>) Representative micrographs showing inhibition of endothelial tube formation. Cells were pretreated with ROS scavengers: ebselen (Ebs.) or N-acetylcysteine (NAC) for 2 h prior to treatment with 1 ng/ml of PCB153 and PCB77. Immunostaining for the endothelial marker CD31 at day 3 showed significant tube formation in the PCB treatment groups compared to vehicle control (0.1% DMSO) that was suppressed by ebselen and NAC treatment. (<b>B</b>) The effect of ROS scavengers on PCB-induced endothelial tube formation was quantified by counting the number of branch points. Co-cultures were established in triplicate and data are expressed as mean ± S.D. between different wells. Measurements were performed from a representative experiment repeated three times in triplicate. (*) PCB treatment significantly different from vehicle control. (**) ROS scavenger treatment significantly different from PCB (<i>P</i><0.05). (<b>C</b>) ID3 siRNA treatments inhibit PCB—induced tube formation in HUVECs co-cultured with fibroblasts at 3 days. Immunostaining for the endothelial cell marker CD31 was used to visualize vessel formation. HUVECs were transfected with ID3 siRNA [50 nM] for 48 h prior to co-culture experiments. Scrambled siRNA (Silencer negative control siRNA, Ambion) was used in all RNAi experiments. The data shown are representative images from an experiment repeated three separate times.</p>", "links"=>[], "tags"=>["cell biology", "Cell processes", "Cell proliferation", "oxidative stress", "physiology", "Cardiovascular physiology", "angiogenesis", "Public and occupational health", "Environmental health", "Vascular medicine", "Vascular diseases", "scavengers", "sirna", "id3", "inhibit", "pcb-induced", "neovascular"], "article_id"=>1126898, "categories"=>["Biological Sciences"], "users"=>["Jayanta K. Das", "Quentin Felty"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0104159.g002", "stats"=>{"downloads"=>0, "page_views"=>20, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_ROS_scavengers_and_siRNA_ID3_inhibit_the_PCB_induced_neovascular_phenotype_/1126898", "title"=>"ROS scavengers and siRNA ID3 inhibit the PCB-induced neovascular phenotype.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-08-04 03:13:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/1620942"], "description"=>"<p>(<b>A</b>) Graph of cell survival from exposure to PCB153 in B27 media determined by MTT assay of endothelial spheroids. Cell survival was determined 24 h after exposure. LC<sub>50</sub>∼100 µM. <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0104159#s3\" target=\"_blank\">Results</a> shown are averages of triplicates ± SD (<10%). (<b>B</b>) Graph of cell survival from exposure to PCB153 in monolayer culture in routine culture media determined by MTT assay. Cell survival was determined 24 h after exposure to PCB153. LC<sub>50</sub>∼45 µM. <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0104159#s3\" target=\"_blank\">Results</a> shown are averages of triplicates ± SD (<10%). (<b>C</b>) PCB153 (60 µM) treatment increased the diameter of endothelial spheroids measured at both days 5 and 10. The hCMEC/D3 cell line was cultured with serum free B27 medium in ultra-low attachment 96 well plates. Spheroid diameter was measured at days 5 and 10. The measurement scale was 100 µm at a magnification of 200×. Error bars represent the mean diameter of 15 spheroids ± SD. Data were analyzed by ANOVA; Tukey HSD test for multiple comparisons. Endothelial spheroid size is the mean diameter of 15 spheroids per treatment group ± SD. <sup>a</sup>p<0.01; <sup>b</sup>p<0.01; <sup>c</sup>p<0.01 vs control within 5 and 10 days as well as <sup>e</sup>p<0.01; <sup>f</sup>p<0.05 vs control between 0 and 5 days and <sup>g</sup>p<0.01; <sup>h</sup>p<0.01 vs control between 0 and 10 days. Representative microphotographs of treatment groups are shown inset.</p>", "links"=>[], "tags"=>["cell biology", "Cell processes", "Cell proliferation", "oxidative stress", "physiology", "Cardiovascular physiology", "angiogenesis", "Public and occupational health", "Environmental health", "Vascular medicine", "Vascular diseases", "endothelial"], "article_id"=>1126900, "categories"=>["Biological Sciences"], "users"=>["Jayanta K. Das", "Quentin Felty"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0104159.g003", "stats"=>{"downloads"=>3, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_PCB153_treatment_increased_the_survival_and_size_of_endothelial_spheroids_/1126900", "title"=>"PCB153 treatment increased the survival and size of endothelial spheroids.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-08-04 03:13:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/1620946"], "description"=>"<p>(<b>A</b>) Representative immunohistochemical analysis of heart tissue using ID3 rabbit monoclonal antibody. ECs from normal tissue did not exhibit detectable amounts of ID3 protein (arrowhead) while (arrows) indicated brown color as highest expression level for ECs with ID3 in vessels from benign and malignant heart tissue. Counter-stained with Hematoxylin QS. Magnification 200×. (<b>B</b>) Scatter plot of immunoreactivity (IR) score vs. ID3 in normal, benign, malignant heart. Spearman's rank correlation test showed a significant correlation between ID3 immunostaining and degree of malignancy with the ID3 IR scores significantly higher in benign compared to normal (p = 0.04) and in malignant compared to normal (p = 0.02). (<b>C</b>) Immunofluorescence staining of ID3 (green) and 8-OHdG (red) in heart tissue. Scale = 20 µm. (<b>D</b>) Scatter plot of immunofluorescence (IF) score vs ID3 & 8-OHdG labeling index (LI) in normal, benign, and malignant heart. IF score represents total score taken as the product of fluorescence intensity score and the cell score. Labeling index (LI) represents the percentage of the immunostained cells per total number of cells. Corresponding graphs to each row of images indicate a higher ID3 & 8-OHdG LI in both malignant and benign tissues compared to normal tissue (p<0.05, Mann-Whitney U-test). (E), Immunofluorescence staining of Id3 (green) and 8-OHdG (red) in main artery tissue. Scale = 20 µm. (F) Scatter plot of IF score vs ID3 & 8-OHdG LI in benign and normal artery tissue. IF score represents total score taken as the product of fluorescence intensity score and the cell score.</p>", "links"=>[], "tags"=>["cell biology", "Cell processes", "Cell proliferation", "oxidative stress", "physiology", "Cardiovascular physiology", "angiogenesis", "Public and occupational health", "Environmental health", "Vascular medicine", "Vascular diseases", "oxidative", "benign", "malignant", "cardiovascular"], "article_id"=>1126903, "categories"=>["Biological Sciences"], "users"=>["Jayanta K. Das", "Quentin Felty"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0104159.g004", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_ID3_protein_and_oxidative_damage_is_highly_expressed_in_benign_and_malignant_cardiovascular_tissues_/1126903", "title"=>"ID3 protein and oxidative damage is highly expressed in benign and malignant cardiovascular tissues.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-08-04 03:13:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/1620949"], "description"=>"<p>(<b>A</b>) PCB153 [5 µM] treatment increased serine phosphorylation of ID3 protein at 3 h. ID3 protein was isolated by immunoprecipitation (IP) technique using magnetic beads and detected by immunoblot. Serine phosphorylation of immuoprecipitated ID3 protein was determined with anti-p-Ser antibody. (<b>B</b>) PCB153 induced ID3 protein was not modulated by aryl hydrocarbon receptor (AhR) at 3 h in hCMEC/D3 cell line. Western blot analysis showed that the AhR ligand-selective antagonist, CH-223191 [10 nM], did not prevent PCB153 induced expression of ID3. (<b>C</b>) PCB153 induced expression of ID3 protein was inhibited by ROS scavenger NAC in hCMEC/D3. (<b>D</b>) PCB153-treated cells in the absence of protein synthesis showed that ID3 protein levels remained higher than control at 3 and 6 h time points. Cells were pretreated with the protein synthesis inhibitor cycloheximide [1 µg/ml] 2 h prior to treatment with PCB153.</p>", "links"=>[], "tags"=>["cell biology", "Cell processes", "Cell proliferation", "oxidative stress", "physiology", "Cardiovascular physiology", "angiogenesis", "Public and occupational health", "Environmental health", "Vascular medicine", "Vascular diseases", "id3", "ahr"], "article_id"=>1126906, "categories"=>["Biological Sciences"], "users"=>["Jayanta K. Das", "Quentin Felty"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0104159.g005", "stats"=>{"downloads"=>2, "page_views"=>20, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_PCB_induced_ID3_expression_is_independent_of_AhR_and_ID3_protein_is_stablized_/1126906", "title"=>"PCB-induced ID3 expression is independent of AhR and ID3 protein is stablized.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-08-04 03:13:24"}

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