TRPM4 Is a Novel Component of the Adhesome Required for Focal Adhesion Disassembly, Migration and Contractility
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{"title"=>"TRPM4 is a novel component of the adhesome required for focal adhesion disassembly, migration and contractility", "type"=>"generic", "authors"=>[{"first_name"=>"Mónica", "last_name"=>"Cáceres", "scopus_author_id"=>"8551903200"}, {"first_name"=>"Liliana", "last_name"=>"Ortiz", "scopus_author_id"=>"56763670000"}, {"first_name"=>"Tatiana", "last_name"=>"Recabarren", "scopus_author_id"=>"56764433200"}, {"first_name"=>"Anibal", "last_name"=>"Romero", "scopus_author_id"=>"56734068500"}, {"first_name"=>"Alicia", "last_name"=>"Colombo", "scopus_author_id"=>"7202937208"}, {"first_name"=>"Elías", "last_name"=>"Leiva-Salcedo", "scopus_author_id"=>"6505855876"}, {"first_name"=>"Diego", "last_name"=>"Varela", "scopus_author_id"=>"16687233600"}, {"first_name"=>"José", "last_name"=>"Rivas", "scopus_author_id"=>"56763579700"}, {"first_name"=>"Ian", "last_name"=>"Silva", "scopus_author_id"=>"56763512200"}, {"first_name"=>"Diego", "last_name"=>"Morales", "scopus_author_id"=>"56763596400"}, {"first_name"=>"Camilo", "last_name"=>"Campusano", "scopus_author_id"=>"56764090700"}, {"first_name"=>"Oscar", "last_name"=>"Almarza", "scopus_author_id"=>"55922220200"}, {"first_name"=>"Felipe", "last_name"=>"Simon", "scopus_author_id"=>"7201952657"}, {"first_name"=>"Hector", "last_name"=>"Toledo", "scopus_author_id"=>"55921584700"}, {"first_name"=>"Kang Sik", "last_name"=>"Park", "scopus_author_id"=>"7408066754"}, {"first_name"=>"James S.", "last_name"=>"Trimmer", "scopus_author_id"=>"7004962489"}, {"first_name"=>"Oscar", "last_name"=>"Cerda", "scopus_author_id"=>"6507734557"}], "year"=>2015, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"605398467", "doi"=>"10.1371/journal.pone.0130540", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "pmid"=>"26110647", "issn"=>"19326203", "scopus"=>"2-s2.0-84938630783", "sgr"=>"84938630783"}, "id"=>"795abc2f-e279-39f6-ae64-27f088be6bdd", "abstract"=>"Cellular migration and contractility are fundamental processes that are regulated by a variety of concerted mechanisms such as cytoskeleton rearrangements, focal adhesion turnover, and Ca2+ oscillations. TRPM4 is a Ca2+-activated non-selective cationic channel (Ca2+-NSCC) that conducts monovalent but not divalent cations. Here, we used a mass spectrometry-based proteomics approach to identify putative TRPM4-associated proteins. Interestingly, the largest group of these proteins has actin cytoskeleton-related functions, and among these nine are specifically annotated as focal adhesion-related proteins. Consistent with these results, we found that TRPM4 localizes to focal adhesions in cells from different cellular lineages. We show that suppression of TRPM4 in MEFs impacts turnover of focal adhesions, serum-induced Ca2+ influx, focal adhesion kinase (FAK) and Rac activities, and results in reduced cellular spreading, migration and contractile behavior. Finally, we demonstrate that the inhibition of TRPM4 activity alters cellular contractility in vivo, affecting cutaneous wound healing. Together, these findings provide the first evidence, to our knowledge, for a TRP channel specifically localized to focal adhesions, where it performs a central role in modulating cellular migration and contractility.", "link"=>"http://www.mendeley.com/research/trpm4-novel-component-adhesome-required-focal-adhesion-disassembly-migration-contractility", "reader_count"=>41, "reader_count_by_academic_status"=>{"Unspecified"=>3, "Professor > Associate Professor"=>1, "Librarian"=>1, "Researcher"=>10, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>10, "Student > Postgraduate"=>2, "Student > Master"=>4, "Other"=>1, "Student > Bachelor"=>5, "Professor"=>2}, "reader_count_by_user_role"=>{"Unspecified"=>3, "Professor > Associate Professor"=>1, "Librarian"=>1, "Researcher"=>10, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>10, "Student > Postgraduate"=>2, "Student > Master"=>4, "Other"=>1, "Student > Bachelor"=>5, "Professor"=>2}, "reader_count_by_subject_area"=>{"Unspecified"=>3, "Engineering"=>1, "Biochemistry, Genetics and Molecular Biology"=>13, "Agricultural and Biological Sciences"=>18, "Medicine and Dentistry"=>2, "Computer Science"=>1, "Immunology and Microbiology"=>2, "Neuroscience"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>2}, "Neuroscience"=>{"Neuroscience"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>2}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>18}, "Computer Science"=>{"Computer Science"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>13}, "Unspecified"=>{"Unspecified"=>3}}, "reader_count_by_country"=>{"Sweden"=>1, "United Kingdom"=>1, "Chile"=>1, "Switzerland"=>1}, "group_count"=>1}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/2149980"], "description"=>"<p>Time courses for normalized fluorescence in MEFs loaded with Fura-2 and treated with DMSO (0.1%v/v) (A) and 10 μM 9-phenanthrol (B). Gray lines correspond to [Ca<sup>2+</sup>]<sub>i</sub> changes in individual MEFs (12 cells, 6 independent experiments for Control; 48 cells, 6 independent experiments for 9-phenanthrol treatment). White circles correspond to the mean trace of [Ca<sup>2+</sup>] signals. TRPM4 activity is required for FAK activation. (C) Time-course activation of FAK upon TRPM4 pharmacological inhibition. Cells were depleted for 4 h and stimulated with 10% v/v serum in presence of DMSO and 10 μM 9-phenanthrol. Cells were fixed at 0, 2, 5, 15, 30 and 60 min post-treatment. Then, the cells were stained with Hoechst (blue) and rabbit mAb anti-pY397-FAK (green). Representative images from three independent experiments of MEFs treated with DMSO and 10 μM 9-phenanthrol are shown. Scale bar correspond to 5 μm. (D) Representative immunoblot from MEFs treated with DMSO and 10 μM 9-phenanthrol is shown. Cells were depleted for 4 h and then, stimulated with 10%v/v serum for 30 min. Upper graph corresponds to the densitometric analyses of 3 independent experiments. Statistical analysis was performed using a Mann Whitney test. (E) Time-course activation of FAK upon TRPM4 silencing. MEF-shRNA<sup>Scramble</sup> and MEF-shRNA<sup>TRPM4</sup> were treated as <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0130540#pone.0130540.g004\" target=\"_blank\">Fig 4C</a>, and stained with Hoechst (blue) and rabbit mAb anti-pY397-FAK (green). tRFP (red) was used as transfection marker. Representative images from three independent experiments are shown. Scale bar correspond to 5 μm. (F) Representative immunoblot from MEFs transfected with shRNA<sup>Scramble</sup> and shRNA<sup>TRPM4</sup> is shown. Cells were treated as <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0130540#pone.0130540.g004\" target=\"_blank\">Fig 4D</a>. Upper graph corresponds to the densitometric analyses of 3 independent experiments. Statistical analysis was performed using a Mann Whitney test.</p>", "links"=>[], "tags"=>["TRPM 4", "cutaneous wound healing", "adhesion", "Focal Adhesion Disassembly", "MEFs impacts turnover", "TRPM 4 localizes", "TRPM 4 activity", "protein", "fak", "Contractility Cellular migration", "contractility"], "article_id"=>1464501, "categories"=>["Uncategorised"], "users"=>["Mónica Cáceres", "Liliana Ortiz", "Tatiana Recabarren", "Anibal Romero", "Alicia Colombo", "Elías Leiva-Salcedo", "Diego Varela", "José Rivas", "Ian Silva", "Diego Morales", "Camilo Campusano", "Oscar Almarza", "Felipe Simon", "Hector Toledo", "Kang-Sik Park", "James S. Trimmer", "Oscar Cerda"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0130540.g004", "stats"=>{"downloads"=>2, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_TRPM4_regulates_intracellular_Ca_2_signaling_and_activation_of_FAK_/1464501", "title"=>"TRPM4 regulates intracellular Ca<sup>2+</sup> signaling and activation of FAK.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-06-25 04:04:15"}
  • {"files"=>["https://ndownloader.figshare.com/files/2150022", "https://ndownloader.figshare.com/files/2150023", "https://ndownloader.figshare.com/files/2150024", "https://ndownloader.figshare.com/files/2150025", "https://ndownloader.figshare.com/files/2150026", "https://ndownloader.figshare.com/files/2150027", "https://ndownloader.figshare.com/files/2150028", "https://ndownloader.figshare.com/files/2150029", "https://ndownloader.figshare.com/files/2150030", "https://ndownloader.figshare.com/files/2150031"], "description"=>"<div><p>Cellular migration and contractility are fundamental processes that are regulated by a variety of concerted mechanisms such as cytoskeleton rearrangements, focal adhesion turnover, and Ca<sup>2+</sup> oscillations. TRPM4 is a Ca<sup>2+</sup>-activated non-selective cationic channel (Ca<sup>2+</sup>-NSCC) that conducts monovalent but not divalent cations. Here, we used a mass spectrometry-based proteomics approach to identify putative TRPM4-associated proteins. Interestingly, the largest group of these proteins has actin cytoskeleton-related functions, and among these nine are specifically annotated as focal adhesion-related proteins. Consistent with these results, we found that TRPM4 localizes to focal adhesions in cells from different cellular lineages. We show that suppression of TRPM4 in MEFs impacts turnover of focal adhesions, serum-induced Ca<sup>2+</sup> influx, focal adhesion kinase (FAK) and Rac activities, and results in reduced cellular spreading, migration and contractile behavior. Finally, we demonstrate that the inhibition of TRPM4 activity alters cellular contractility <i>in vivo</i>, affecting cutaneous wound healing. Together, these findings provide the first evidence, to our knowledge, for a TRP channel specifically localized to focal adhesions, where it performs a central role in modulating cellular migration and contractility.</p></div>", "links"=>[], "tags"=>["TRPM 4", "cutaneous wound healing", "adhesion", "Focal Adhesion Disassembly", "MEFs impacts turnover", "TRPM 4 localizes", "TRPM 4 activity", "protein", "fak", "Contractility Cellular migration", "contractility"], "article_id"=>1464532, "categories"=>["Uncategorised"], "users"=>["Mónica Cáceres", "Liliana Ortiz", "Tatiana Recabarren", "Anibal Romero", "Alicia Colombo", "Elías Leiva-Salcedo", "Diego Varela", "José Rivas", "Ian Silva", "Diego Morales", "Camilo Campusano", "Oscar Almarza", "Felipe Simon", "Hector Toledo", "Kang-Sik Park", "James S. Trimmer", "Oscar Cerda"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0130540.s001", "https://dx.doi.org/10.1371/journal.pone.0130540.s002", "https://dx.doi.org/10.1371/journal.pone.0130540.s003", "https://dx.doi.org/10.1371/journal.pone.0130540.s004", "https://dx.doi.org/10.1371/journal.pone.0130540.s005", "https://dx.doi.org/10.1371/journal.pone.0130540.s006", "https://dx.doi.org/10.1371/journal.pone.0130540.s007", "https://dx.doi.org/10.1371/journal.pone.0130540.s008", "https://dx.doi.org/10.1371/journal.pone.0130540.s009", "https://dx.doi.org/10.1371/journal.pone.0130540.s010"], "stats"=>{"downloads"=>17, "page_views"=>17, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_TRPM4_Is_a_Novel_Component_of_the_Adhesome_Required_for_Focal_Adhesion_Disassembly_Migration_and_Contractility_/1464532", "title"=>"TRPM4 Is a Novel Component of the Adhesome Required for Focal Adhesion Disassembly, Migration and Contractility", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2015-06-25 04:04:15"}
  • {"files"=>["https://ndownloader.figshare.com/files/2149976"], "description"=>"<p>A) Representative immunoblot from MEFs subjected to shRNA-mediated TRPM4 knock down. Membranes were incubated with mouse anti-TRPM4 mAb, and anti-Grp75 mAb as a loading control. B) Graph of the densitometric analyses of three independent immunoblot experiments. Statistical analysis was performed using a Mann Whitney test. C) Immunofluorescence labeling of MEFs transfected with shRNA<sup>Scramble</sup>, and shRNA<sup>TRPM4</sup>. Cells were labeled with Hoechst (blue), and mouse anti-TRPM4 mAb (green); tRFP (red) was used as transfection marker. Scale bar corresponds to 10 μm. D) Immunofluorescence labeling of MEFs transfected with shRNA<sup>Scramble</sup> and shRNA<sup>TRPM4</sup>. Cells were labeled with Hoechst (blue), and mouse anti-vinculin mAb (green); tRFP (red) was used as transfection marker. Scale bar corresponds to 10 μm. Quantification of FA number (E) and areas (F) from the shRNA-transfected cells (n = 15 cells for shRNA<sup>Scramble</sup> and n = 15 cells for shRNA<sup>TRPM4</sup> from 7 independent experiments). G) Immunofluorescence labeling of MEFs treated with DMSO (0.1% v/v) and 10 μM 9-phenanthrol. Cells were labeled with Hoechst (blue) and mouse anti-vinculin mAb (green). Scale bar corresponds to 10 μm. Graphs of FA number (H) and areas (I) are shown (20 cells per condition, n = 3, p<0.05). The number and areas of the FAs were analyzed using NIH/ImageJ software. The graphs correspond to mean ± standard deviation. Statistical analysis was performed using a Mann-Whitney test.</p>", "links"=>[], "tags"=>["TRPM 4", "cutaneous wound healing", "adhesion", "Focal Adhesion Disassembly", "MEFs impacts turnover", "TRPM 4 localizes", "TRPM 4 activity", "protein", "fak", "Contractility Cellular migration", "contractility"], "article_id"=>1464497, "categories"=>["Uncategorised"], "users"=>["Mónica Cáceres", "Liliana Ortiz", "Tatiana Recabarren", "Anibal Romero", "Alicia Colombo", "Elías Leiva-Salcedo", "Diego Varela", "José Rivas", "Ian Silva", "Diego Morales", "Camilo Campusano", "Oscar Almarza", "Felipe Simon", "Hector Toledo", "Kang-Sik Park", "James S. Trimmer", "Oscar Cerda"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0130540.g002", "stats"=>{"downloads"=>0, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_TRPM4_regulates_the_number_and_size_of_focal_adhesions_/1464497", "title"=>"TRPM4 regulates the number and size of focal adhesions.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-06-25 04:04:15"}
  • {"files"=>["https://ndownloader.figshare.com/files/2149974"], "description"=>"<p>A) Classification of putative TRPM4-associated proteins identified by LC-MS/MS. B) FA-related proteins identified as putative TRPM4 interacting proteins. C) TRPM4 localizes to FAs in MEFs. Cells were labeled with Hoechst (blue), mouse anti-TRPM4 mAb (green), and tRFP (red). D) Magnification of the section from C. E) TRPM4 (green) colocalizes with Focal Adhesion Kinase (FAK, magenta) in MEFs. F) Amplification of the region marked in E. G) Biochemical isolation of FA complexes (FA) from MEFs. The cell body fraction is labeled as CB. TRPM4 localizes to FAs in mouse pancreatic (H) and skin fibroblasts (I); Human Umbilical Vein Endothelial Cells (HUVEC) (J) and astrocytes (K). Scale bar: 5 μm.</p>", "links"=>[], "tags"=>["TRPM 4", "cutaneous wound healing", "adhesion", "Focal Adhesion Disassembly", "MEFs impacts turnover", "TRPM 4 localizes", "TRPM 4 activity", "protein", "fak", "Contractility Cellular migration", "contractility"], "article_id"=>1464495, "categories"=>["Uncategorised"], "users"=>["Mónica Cáceres", "Liliana Ortiz", "Tatiana Recabarren", "Anibal Romero", "Alicia Colombo", "Elías Leiva-Salcedo", "Diego Varela", "José Rivas", "Ian Silva", "Diego Morales", "Camilo Campusano", "Oscar Almarza", "Felipe Simon", "Hector Toledo", "Kang-Sik Park", "James S. Trimmer", "Oscar Cerda"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0130540.g001", "stats"=>{"downloads"=>0, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_TRPM4_localizes_to_focal_adhesions_/1464495", "title"=>"TRPM4 localizes to focal adhesions.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-06-25 04:04:15"}
  • {"files"=>["https://ndownloader.figshare.com/files/2149983"], "description"=>"<p>A) MEFs were incubated with DMSO (0.1% v/v final) vehicle or 10 μM 9-phenanthrol for 16 h. F-actin was labeled with the F-actin stain Alexa 555 phalloidin (red) and Hoechst (blue). B) Quantified results from A. The percentage of wound closure is expressed as mean ± standard deviation; s.d. (n = 3, p<0.05). *, significant difference (p<0.05) versus DMSO controls. Statistical analysis was performed using a Mann-Whitney test. C) Graph from Transwell Boyden chamber migration assays of MEFs transfected with shRNA<sup>Scramble</sup> and shRNA<sup>TRPM4</sup>. Cells were stimulated with 10% v/v serum for 16 h. The bars represent the mean ± s.d. (n = 3; p<0.05 compared to control). *, significant difference (p<0.05) versus shRNA<sup>Scramble</sup> controls. Statistical analysis was performed using a two-way ANOVA test. D) Graph from Transwell Boyden chamber migration assays of MEFs transfected with shRNA<sup>Scramble</sup> and shRNA<sup>TRPM4</sup> and coexpressing Rac1(Q61L). Cells were stimulated with 10% v/v serum for 16 h. The bars represent the mean ± s.d. (n = 3; p<0.05 compared to control). *, significant difference (p<0.05) versus shRNA<sup>Scramble</sup>/Mock controls. **, significant difference (p<0.05) versus shRNA<sup>TRPM4</sup>/Mock controls. Statistical analysis was performed using a two-way ANOVA test.</p>", "links"=>[], "tags"=>["TRPM 4", "cutaneous wound healing", "adhesion", "Focal Adhesion Disassembly", "MEFs impacts turnover", "TRPM 4 localizes", "TRPM 4 activity", "protein", "fak", "Contractility Cellular migration", "contractility"], "article_id"=>1464504, "categories"=>["Uncategorised"], "users"=>["Mónica Cáceres", "Liliana Ortiz", "Tatiana Recabarren", "Anibal Romero", "Alicia Colombo", "Elías Leiva-Salcedo", "Diego Varela", "José Rivas", "Ian Silva", "Diego Morales", "Camilo Campusano", "Oscar Almarza", "Felipe Simon", "Hector Toledo", "Kang-Sik Park", "James S. Trimmer", "Oscar Cerda"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0130540.g006", "stats"=>{"downloads"=>2, "page_views"=>19, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_TRPM4_controls_cellular_migration_via_Rac1_GTPase_activity_/1464504", "title"=>"TRPM4 controls cellular migration <i>via</i> Rac1 GTPase activity.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-06-25 04:04:15"}
  • {"files"=>["https://ndownloader.figshare.com/files/2149981"], "description"=>"<p>A) Cells were incubated with DMSO and 10 μM 9-phenanthrol for 40 min on Collagen-coated coverslips. Cells were fixed and labeled with the F-actin stain Alexa 488 phalloidin (green) and Hoechst (blue). (Scale bar: 10 μm). B) Magnification of cells in A. (Scale bar: 5 μm). C) Graph shows the percentage of cells with lamellipodia-like pattern. Data are from 8 independent coverslips from 3 independent experiments. *, significant difference (p<0.05) versus DMSO controls. Statistical analysis was performed using a Mann-Whitney test. D) Rac GTPase activity of MEF-shRNA<sup>Scramble</sup> and MEF-shRNA<sup>TRPM4</sup> determined using a G-ELISA commercial kit. Rac GTPase activity was stimulated with 5% v/v serum for 30 minutes. *, significant difference (p<0.05) versus shRNA<sup>Scramble</sup> controls. Statistical analysis was performed using a two-way ANOVA test. E) Expression of Rac1(Q61L) rescues the loss of lamellipodial formation caused by the depletion of TRPM4. F-actin was labeled with the F-actin stain Alexa 488 phalloidin (green). RFP (red) corresponds to shRNA-transfected cells. (Scale bar: 5 μm). F) Quantification of the percentages of cells with lamellipodia-like phenotype from experiments showed in E. The graph corresponds to data collected from 3 independent experiments (2 coverslips per condition). *, significant difference (p<0.05) versus shRNA<sup>Scramble</sup>/Mock controls. **, significant difference (p<0.05) versus shRNA<sup>TRPM4</sup>/Mock. Statistical analysis was performed using a two-way ANOVA test.</p>", "links"=>[], "tags"=>["TRPM 4", "cutaneous wound healing", "adhesion", "Focal Adhesion Disassembly", "MEFs impacts turnover", "TRPM 4 localizes", "TRPM 4 activity", "protein", "fak", "Contractility Cellular migration", "contractility"], "article_id"=>1464502, "categories"=>["Uncategorised"], "users"=>["Mónica Cáceres", "Liliana Ortiz", "Tatiana Recabarren", "Anibal Romero", "Alicia Colombo", "Elías Leiva-Salcedo", "Diego Varela", "José Rivas", "Ian Silva", "Diego Morales", "Camilo Campusano", "Oscar Almarza", "Felipe Simon", "Hector Toledo", "Kang-Sik Park", "James S. Trimmer", "Oscar Cerda"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0130540.g005", "stats"=>{"downloads"=>2, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_TRPM4_channels_regulate_Rac_GTPase_activity_/1464502", "title"=>"TRPM4 channels regulate Rac GTPase activity.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-06-25 04:04:15"}
  • {"files"=>["https://ndownloader.figshare.com/files/2149978"], "description"=>"<p>(A) Representative time-lapse imaging from MEFs coexpressing shRNA<sup>Scramble</sup> or shRNA<sup>TRPM4</sup> with EGFP-Paxillin. Assembling FAs are marked with arrows. Arrowheads mark FAs undergoing turnover. Scale bar: 5 μm. Quantification of assembly (B) and disassembly rates (C) of FAs from live-cell time-lapse recordings of EGFP-Paxillin transfected MEFs. Statistical analysis was performed using a Mann-Whitney test from 7 independent experiments.</p>", "links"=>[], "tags"=>["TRPM 4", "cutaneous wound healing", "adhesion", "Focal Adhesion Disassembly", "MEFs impacts turnover", "TRPM 4 localizes", "TRPM 4 activity", "protein", "fak", "Contractility Cellular migration", "contractility"], "article_id"=>1464498, "categories"=>["Uncategorised"], "users"=>["Mónica Cáceres", "Liliana Ortiz", "Tatiana Recabarren", "Anibal Romero", "Alicia Colombo", "Elías Leiva-Salcedo", "Diego Varela", "José Rivas", "Ian Silva", "Diego Morales", "Camilo Campusano", "Oscar Almarza", "Felipe Simon", "Hector Toledo", "Kang-Sik Park", "James S. Trimmer", "Oscar Cerda"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0130540.g003", "stats"=>{"downloads"=>0, "page_views"=>19, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_TRPM4_regulates_FA_turnover_/1464498", "title"=>"TRPM4 regulates FA turnover.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-06-25 04:04:15"}
  • {"files"=>["https://ndownloader.figshare.com/files/2149990"], "description"=>"<p>A) Three-dimensional (3D) invasion assay of TREx293-TRPM4 cells. TRPM4 expression was induced by adding 1 μg/mL Tetracycline to the media (n = 3, p<0.05 compared to control). *, significant difference (p<0.05) versus non-stimulated cell control. Statistical analysis was performed using a Mann-Whitney test. B) Fibroblasts migration from skin grafts (E) treated with DMSO or 20 μM 9-phenanthrol. Grafts were fixed 5 days after explant and labeled with Hoechst (blue). Scale bar: 1 mm. C) Quantification of the experiment shown in (B). The data correspond to cell counts from 3 independent experiments (13 and 12 explants for DMSO and 9-phenanthrol treatments, respectively). *, significant difference (p<0.05) versus DMSO controls. Statistical analysis was performed using a Mann-Whitney test. D) Three dimensional contraction assay of MEFs transfected with shRNA<sup>Scramble</sup> and shRNA<sup>TRPM4</sup>. Contraction was induced by incubating the immersed cells with 10% v/v serum for 48 h. The upper graph represents the collected data for 4 independent assays. *, significant difference (p<0.05) versus shRNA<sup>Scramble</sup> controls. **, significant difference (p<0.05) versus shRNA<sup>TRPM4</sup>. Statistical analysis was performed using a two-way ANOVA test. E) Frames (t = 0 and 14 min) from time lapse of untreated, DMSO and 9-phenanthrol treated wounds in zebrafish tails. Scale bar: 1 mm. F) Quantification of the wound closure experiments from (E) (n = 10 larvae per condition). Statistical analysis was performed using a two-way ANOVA test. G) Excisional cutaneous wounds were created using a 3 mm biopsy punch. Images from the time course of wound closure in the presence of DMSO (control) and 9-phenanthrol (n = 5 mice). Scale bar: 1.5 mm. H) Wound closure was monitored measuring the area of the wound on the indicated days post-wounding. Statistical analysis was performed using a two-way ANOVA test. I) Images of skin wounds at 3 days post-wounding. Bottom panels show magnifications of the areas marked in the upper panels. Arrowheads mark the epithelial tissue. Scale bar: 50 μm. J) Images of wounds at 5 days post-wounding. The dashed lines indicate the limit of the wound area. Scale bar: 500 μm.</p>", "links"=>[], "tags"=>["TRPM 4", "cutaneous wound healing", "adhesion", "Focal Adhesion Disassembly", "MEFs impacts turnover", "TRPM 4 localizes", "TRPM 4 activity", "protein", "fak", "Contractility Cellular migration", "contractility"], "article_id"=>1464511, "categories"=>["Uncategorised"], "users"=>["Mónica Cáceres", "Liliana Ortiz", "Tatiana Recabarren", "Anibal Romero", "Alicia Colombo", "Elías Leiva-Salcedo", "Diego Varela", "José Rivas", "Ian Silva", "Diego Morales", "Camilo Campusano", "Oscar Almarza", "Felipe Simon", "Hector Toledo", "Kang-Sik Park", "James S. Trimmer", "Oscar Cerda"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0130540.g007", "stats"=>{"downloads"=>2, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_TRPM4_promotes_cellular_contractility_and_wound_healing_/1464511", "title"=>"TRPM4 promotes cellular contractility and wound healing.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-06-25 04:04:15"}

PMC Usage Stats | Further Information

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  • {"unique-ip"=>"15", "full-text"=>"14", "pdf"=>"4", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"11", "supp-data"=>"8", "cited-by"=>"0", "year"=>"2019", "month"=>"5"}
  • {"unique-ip"=>"15", "full-text"=>"17", "pdf"=>"5", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"1", "year"=>"2019", "month"=>"8"}
  • {"unique-ip"=>"15", "full-text"=>"14", "pdf"=>"9", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"9"}
  • {"unique-ip"=>"17", "full-text"=>"23", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"10"}

Relative Metric

{"start_date"=>"2015-01-01T00:00:00Z", "end_date"=>"2015-12-31T00:00:00Z", "subject_areas"=>[]}
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