Detachment of Breast Tumor Cells Induces Rapid Secretion of Exosomes Which Subsequently Mediate Cellular Adhesion and Spreading
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{"title"=>"Detachment of breast tumor cells induces rapid secretion of exosomes which subsequently mediate cellular adhesion and spreading", "type"=>"journal", "authors"=>[{"first_name"=>"Rainelli B.", "last_name"=>"Koumangoye", "scopus_author_id"=>"36617180000"}, {"first_name"=>"Amos M.", "last_name"=>"Sakwe", "scopus_author_id"=>"6603131076"}, {"first_name"=>"J. Shawn", "last_name"=>"Goodwin", "scopus_author_id"=>"7201951448"}, {"first_name"=>"Tina", "last_name"=>"Patel", "scopus_author_id"=>"51663855100"}, {"first_name"=>"Josiah", "last_name"=>"Ochieng", "scopus_author_id"=>"7003905679"}], "year"=>2011, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "scopus"=>"2-s2.0-80052446161", "sgr"=>"80052446161", "pui"=>"362504708", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "pmid"=>"21915303", "doi"=>"10.1371/journal.pone.0024234"}, "id"=>"24e5aae9-3795-33b2-9602-6fa8bc42aa41", "abstract"=>"Exosomes are nano-vesicles secreted by a wide range of mammalian cell types. These vesicles are abundant in serum and other extracellular fluids and contain a large repertoire of proteins, mRNA and microRNA. Exosomes have been implicated in cell to cell communication, the transfer of infectious agents, and neurodegenerative diseases as well as tumor progression. However, the precise mechanisms by which they are internalized and/or secreted remain poorly understood. In order to follow their release and uptake in breast tumor cells in real time, cell-derived exosomes were tagged with green fluorescent protein (GFP)-CD63 while human serum exosomes were rhodamine isothiocynate-labeled. We show that detachment of adherent cells from various substrata induces a rapid and substantial secretion of exosomes, which then concentrate on the cell surfaces and mediate adhesion to various extracellular matrix proteins. We also demonstrate that disruption of lipid rafts with methyl-beta-cyclodextrin (MbetaCD) inhibits the internalization of exosomes and that annexins are essential for the exosomal uptake mechanisms. Taken together, these data suggest that cellular detachment is accompanied by significant release of exosomes while cellular adhesion and spreading are enhanced by rapid uptake and disposition of exosomes on the cell surface.", "link"=>"http://www.mendeley.com/research/detachment-breast-tumor-cells-induces-rapid-secretion-exosomes-subsequently-mediate-cellular-adhesio", "reader_count"=>143, "reader_count_by_academic_status"=>{"Unspecified"=>5, "Professor > Associate Professor"=>9, "Researcher"=>34, "Student > Doctoral Student"=>9, "Student > Ph. D. Student"=>38, "Student > Postgraduate"=>4, "Other"=>4, "Student > Master"=>16, "Student > Bachelor"=>17, "Lecturer"=>2, "Lecturer > Senior Lecturer"=>1, "Professor"=>4}, "reader_count_by_user_role"=>{"Unspecified"=>5, "Professor > Associate Professor"=>9, "Researcher"=>34, "Student > Doctoral Student"=>9, "Student > Ph. D. Student"=>38, "Student > Postgraduate"=>4, "Other"=>4, "Student > Master"=>16, "Student > Bachelor"=>17, "Lecturer"=>2, "Lecturer > Senior Lecturer"=>1, "Professor"=>4}, "reader_count_by_subject_area"=>{"Unspecified"=>7, "Agricultural and Biological Sciences"=>73, "Arts and Humanities"=>1, "Philosophy"=>1, "Veterinary Science and Veterinary Medicine"=>1, "Chemical Engineering"=>1, "Chemistry"=>8, "Engineering"=>5, "Biochemistry, Genetics and Molecular Biology"=>21, "Materials Science"=>1, "Medicine and Dentistry"=>19, "Neuroscience"=>2, "Sports and Recreations"=>1, "Pharmacology, Toxicology and Pharmaceutical Science"=>2}, "reader_count_by_subdiscipline"=>{"Materials Science"=>{"Materials Science"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>19}, "Sports and Recreations"=>{"Sports and Recreations"=>1}, "Unspecified"=>{"Unspecified"=>7}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>2}, "Chemical Engineering"=>{"Chemical Engineering"=>1}, "Arts and Humanities"=>{"Arts and Humanities"=>1}, "Engineering"=>{"Engineering"=>5}, "Chemistry"=>{"Chemistry"=>8}, "Neuroscience"=>{"Neuroscience"=>2}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>73}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>21}, "Philosophy"=>{"Philosophy"=>1}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>1}}, "reader_count_by_country"=>{"Sweden"=>1, "Hungary"=>1, "Belgium"=>1, "United States"=>1, "China"=>1, "United Kingdom"=>1, "Chile"=>1, "France"=>1, "Spain"=>1}, "group_count"=>8}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/739812"], "description"=>"<p>BT-549 stably transfected with either GFP-AnxA6 (panel A) or GFP-AnxA2 (panel B), were plated on glass coverslips in culture medium. After 24 h, the cells were washed 3X with PBS, loaded or depleted of cholesterol. Fluorescence recover after photo-bleaching (FRAP) was performed on a Nikon A1R laser scanning confocal microscopy as detailed in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0024234#s2\" target=\"_blank\">Materials and Methods</a>. FRAP readings were done on cells in PBS, pH 7.4 and at room temperature.</p>", "links"=>[], "tags"=>["oncology", "women's health", "Biochemistry"], "article_id"=>410189, "categories"=>["Cancer", "Biochemistry", "Mathematics"], "users"=>["Rainelli B. Koumangoye", "Amos M. Sakwe", "J. Shawn Goodwin", "Tina Patel", "Josiah Ochieng"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0024234.g009", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Fluorescence_recovery_after_photo_bleaching_/410189", "title"=>"Fluorescence recovery after photo-bleaching.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-09-06 00:03:09"}
  • {"files"=>["https://ndownloader.figshare.com/files/739322"], "description"=>"<p>BT-549 cells were grown on glass cover-slips for 24 hours. Cells were then washed twice with HBSS, and treated with different concentrations of methyl-beta-cyclodextrin (MβCD) (0–10 mM) for 30 minutes. Subsequently, cells were incubated with 10 µg/ml GFP-CD63 labeled exosomes in HBSS containing 0.5% BSA and 1 mM Ca<sup>2+</sup>/Mg<sup>2+</sup> for 30 minutes at 37°C. In panel A, the untreated (control) and cells pre-incubated with 10 mM of MβCD were washed in ice-cold HBSS and fixed with paraformaldehyde and cover-slips mounted with Prolong Gold and visualized by confocal microscopy. Images were captured using a Nikon TE2000. Bar is 20 µm. In panel B, the cells were also processed for flow cytometry as indicated. Unstained control cells (black line) were included. In panel C, BT-549 cells grown on MatTek glass bottom culture dishes were incubated in HBSS 0.5% BSA 1 mM Ca<sup>2+</sup> 1 mM Mg<sup>2+</sup> on ice for 10 minutes. The cells were then treated with 10 µg/mL of alexa fluor Tf (left panel) or CTXB (right panel) and GFP-labeled exosomes on ice for 10 minutes again. The trafficking of GFP labeled particles and colocalization with either Tf or CTXB (white arrows) were tracked live on the Nikon TE2000 for 10 minutes. Representative time-lapse images are shown.</p>", "links"=>[], "tags"=>["exosomal", "uptake", "carcinoma"], "article_id"=>409694, "categories"=>["Cancer", "Biochemistry", "Mathematics"], "users"=>["Rainelli B. Koumangoye", "Amos M. Sakwe", "J. Shawn Goodwin", "Tina Patel", "Josiah Ochieng"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0024234.g006", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_exosomal_uptake_route_in_breast_carcinoma_cells_/409694", "title"=>"The exosomal uptake route in breast carcinoma cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-09-06 02:41:34"}
  • {"files"=>["https://ndownloader.figshare.com/files/739460"], "description"=>"<p>BT-549 cells gown on glass cover slips were incubated with purified GFP-CD63 exosomes (green) for the indicated time points. At each time point the cells were fixed with PFA and stained with antibodies to the early, late and recycling endosomes followed by TRITC conjugated secondary antibodies (red). The first row shows co-localization of GFP-exosomes with EEA1 (early endosomes); Second row shows colocalization with CD71 (recycling endosomes); and the third row shows colocalization with LAMP1 (late endosomes). Bar is 10 µm.</p>", "links"=>[], "tags"=>["gfp-exosomes", "carcinoma"], "article_id"=>409833, "categories"=>["Cancer", "Biochemistry", "Mathematics"], "users"=>["Rainelli B. Koumangoye", "Amos M. Sakwe", "J. Shawn Goodwin", "Tina Patel", "Josiah Ochieng"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0024234.g007", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Trafficking_of_GFP_exosomes_in_breast_carcinoma_cells_/409833", "title"=>"Trafficking of GFP-exosomes in breast carcinoma cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-09-06 02:43:53"}
  • {"files"=>["https://ndownloader.figshare.com/files/739659"], "description"=>"<p>Parental BT-549, BT-A2-sh and BT-A6-sh were grown on glass coverslips or 10 cm dishes and incubated with GFP-CD63 exosomes and analyzed by (A) confocal microscopy or (B) Flow Cytometry. In panels C and D, BT549 cells were grown on glass cover-slips, serum starved for 24 hours washed and stimulated without (control) and with 100 µg/mL exosomes in HBSS 0.5% BSA 1 mM Ca<sup>2+</sup> 1 mM Mg<sup>2+</sup> for 5 and 15 minutes at 37°C and fixed with PFA. Cells were then treated with rhodamine phalloidin for actin staining followed by antibodies to AnxA2 (C) and AnxA6 (D). Images were acquired with a Nikon TE2000 confocal microscope. Bar is 20 µm.</p>", "links"=>[], "tags"=>["uptake", "disposition", "exosomes"], "article_id"=>410037, "categories"=>["Cancer", "Biochemistry", "Mathematics"], "users"=>["Rainelli B. Koumangoye", "Amos M. Sakwe", "J. Shawn Goodwin", "Tina Patel", "Josiah Ochieng"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0024234.g008", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Annexin_mediated_uptake_and_disposition_of_exosomes_on_the_cell_surface_/410037", "title"=>"Annexin-mediated uptake and disposition of exosomes on the cell surface.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-09-06 00:00:37"}
  • {"files"=>["https://ndownloader.figshare.com/files/739930"], "description"=>"<p>BT-549 cells gown on glass cover slips were incubated without or with purified GFP-CD63 exosomes (green) for 2 h. The cells were then fixed with PFA and stained with antibodies to AnxA2 (panel A) or AnxA6 (panel B) followed by TRITC conjugated secondary antibodies (red). Bar is 10 µm.</p>", "links"=>[], "tags"=>["anxa6", "anxa2", "endosomal", "compartments", "exosomal", "uptake", "bt-549"], "article_id"=>410302, "categories"=>["Cancer", "Biochemistry", "Mathematics"], "users"=>["Rainelli B. Koumangoye", "Amos M. Sakwe", "J. Shawn Goodwin", "Tina Patel", "Josiah Ochieng"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0024234.g010", "stats"=>{"downloads"=>1, "page_views"=>18, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Trafficking_of_AnxA6_but_not_AnxA2_to_the_endosomal_compartments_as_a_function_of_exosomal_uptake_by_BT_549_cells_/410302", "title"=>"Trafficking of AnxA6 but not AnxA2 to the endosomal compartments as a function of exosomal uptake by BT-549 cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-09-06 00:05:02"}
  • {"files"=>["https://ndownloader.figshare.com/files/738875"], "description"=>"<p>A-B) BT-CD63 cells were maintained in adherent (panel A) or non-adherent (panel B) modes as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0024234#s2\" target=\"_blank\">materials and methods</a> and examined by confocal microscopy Note that the EDTA-induced detachment of BT-CD63 led to the accumulation of the GFP-CD63 labeled exosomes on the cell surface. C-D), parental BT-549 cells in adherent cultures or in anchorage-independent cultures on polyHEMA coated plates were incubated with conditioned medium containing GFP-CD63 vesicles for 3 h. Cells were thereafter processed for microcopy as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0024234#s2\" target=\"_blank\">Materials and Methods</a>. Bar is 10 µm.</p>", "links"=>[], "tags"=>["gfp-cd63", "containing", "exosomes", "adherent", "non-adherent", "bt-cd63"], "article_id"=>409245, "categories"=>["Cancer", "Biochemistry", "Mathematics"], "users"=>["Rainelli B. Koumangoye", "Amos M. Sakwe", "J. Shawn Goodwin", "Tina Patel", "Josiah Ochieng"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0024234.g003", "stats"=>{"downloads"=>4, "page_views"=>248, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Distribution_of_GFP_CD63_containing_exosomes_in_adherent_and_non_adherent_BT_CD63_cells_/409245", "title"=>"Distribution of GFP-CD63 containing exosomes in adherent and non-adherent BT-CD63 cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-09-06 02:34:05"}
  • {"files"=>["https://ndownloader.figshare.com/files/738734"], "description"=>"<p>BT-CD63 cells or GFP expressing control cells were plated on MatTek glass dishes for 24 hours. Prior to live-cell imaging cells were washed 2 times in PBS. Cells were then treated with EDTA (2 mM) and the trafficking and release of GFP-labeled exosomes was monitored by time-lapse imaging for up to 10 minutes. Upper panels are representative individual frames from time-lapse acquired images of GFP expressing control cells. Lower panels are representative individual frames tracking the release of the GFP-tagged CD63 vesicles. The white arrow shows a strongly adhered cell that was not detached even after 10 minutes following EDTA addition. The yellow arrows indicate stream of GFP-tagged CD63 vesicles that are being released (see <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0024234#pone.0024234.s002\" target=\"_blank\">movie S1</a> and <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0024234#pone.0024234.s003\" target=\"_blank\">S2</a> in supplemental materials). Bar is 20 µm.</p>", "links"=>[], "tags"=>["cellular", "detachment-induced", "exosomes", "bt-cd63"], "article_id"=>409102, "categories"=>["Cancer", "Biochemistry", "Mathematics"], "users"=>["Rainelli B. Koumangoye", "Amos M. Sakwe", "J. Shawn Goodwin", "Tina Patel", "Josiah Ochieng"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0024234.g002", "stats"=>{"downloads"=>2, "page_views"=>54, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Visualization_in_real_time_of_cellular_detachment_induced_rapid_release_of_exosomes_from_BT_CD63_cells_/409102", "title"=>"Visualization in real time of cellular detachment-induced rapid release of exosomes from BT-CD63 cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-09-06 02:31:42"}
  • {"files"=>["https://ndownloader.figshare.com/files/739205"], "description"=>"<p>In panel A, BT-CD63 cells were plated in 35 mm with glass bottom dishes and allowed to adhere for at least 24 hours in complete DMEM/F-12 medium. The culture medium was replaced with fresh medium containing 30 µM BAPTA-AM and allowed to incubate for another 7 hours. The dish was then placed on microscope stage (Nikon A1R) and the live cells photographed (t = 0) and then 2 mM EDTA added. After 5 and 10 minutes of EDTA addition, images were again taken of the cells. Bar is 20 µm. In panel B, BT-549-Gal-3 cells in control wells (untreated) and in BAPTA-AM treated wells (in the absence and presence of purified cellular exosomes) were allowed to adhere to Fetuin-A coated wells, fixed, stained and photographed as described above. Bar is 40 µm. In panel C, the experiments in B were repeated with a different cell line (MDA-MB-231). At the end of the incubation, the non-adhered cells were washed off and fresh medium containing a 1∶10 dilution of Alamar Blue added to the wells, incubated for another 1 h and cell number determined by fluorescence spectroscopy.</p>", "links"=>[], "tags"=>["secreted", "cells", "detachment", "adhesion"], "article_id"=>409579, "categories"=>["Cancer", "Biochemistry", "Mathematics"], "users"=>["Rainelli B. Koumangoye", "Amos M. Sakwe", "J. Shawn Goodwin", "Tina Patel", "Josiah Ochieng"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0024234.g005", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Exosomes_secreted_by_tumor_cells_during_detachment_are_required_for_subsequent_adhesion_and_spreading_/409579", "title"=>"Exosomes secreted by tumor cells during detachment are required for subsequent adhesion and spreading.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-09-06 02:39:39"}
  • {"files"=>["https://ndownloader.figshare.com/files/372869", "https://ndownloader.figshare.com/files/372929", "https://ndownloader.figshare.com/files/372986"], "description"=>"<div><p>Exosomes are nano-vesicles secreted by a wide range of mammalian cell types. These vesicles are abundant in serum and other extracellular fluids and contain a large repertoire of proteins, mRNA and microRNA. Exosomes have been implicated in cell to cell communication, the transfer of infectious agents, and neurodegenerative diseases as well as tumor progression. However, the precise mechanisms by which they are internalized and/or secreted remain poorly understood. In order to follow their release and uptake in breast tumor cells in real time, cell-derived exosomes were tagged with green fluorescent protein (GFP)-CD63 while human serum exosomes were rhodamine isothiocynate-labeled. We show that detachment of adherent cells from various substrata induces a rapid and substantial secretion of exosomes, which then concentrate on the cell surfaces and mediate adhesion to various extracellular matrix proteins. We also demonstrate that disruption of lipid rafts with methyl-beta-cyclodextrin (MβCD) inhibits the internalization of exosomes and that annexins are essential for the exosomal uptake mechanisms. Taken together, these data suggest that cellular detachment is accompanied by significant release of exosomes while cellular adhesion and spreading are enhanced by rapid uptake and disposition of exosomes on the cell surface.</p> </div>", "links"=>[], "tags"=>["detachment", "cells", "induces", "secretion", "exosomes", "subsequently", "mediate", "cellular", "adhesion", "spreading"], "article_id"=>133637, "categories"=>["Cancer", "Biochemistry", "Mathematics"], "users"=>["Rainelli B. Koumangoye", "Amos M. Sakwe", "J. Shawn Goodwin", "Tina Patel", "Josiah Ochieng"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0024234.s001", "https://dx.doi.org/10.1371/journal.pone.0024234.s002", "https://dx.doi.org/10.1371/journal.pone.0024234.s003"], "stats"=>{"downloads"=>3, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Detachment_of_Breast_Tumor_Cells_Induces_Rapid_Secretion_of_Exosomes_Which_Subsequently_Mediate_Cellular_Adhesion_and_Spreading/133637", "title"=>"Detachment of Breast Tumor Cells Induces Rapid Secretion of Exosomes Which Subsequently Mediate Cellular Adhesion and Spreading", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2011-09-06 01:00:37"}
  • {"files"=>["https://ndownloader.figshare.com/files/738627"], "description"=>"<p>(A) BT-549 cells stably expressing GFP-CD63 were grown on glass cover-slips for 24 hours and the intracellular localization of the CD63-associated vesicles visualized by confocal microscopy. Bar is 10 µm. (B) Over-expression of GFP-CD63. Total cell lysates from control BT-549 and BT-549 expressing GFP-CD63 (BT-CD63) cells were analyzed by western blotting and probed with antibodies against CD63. (C) Exosomes purified from the spent media of BT-CD63 cells were layered on top of a sucrose step gradient and centrifuged at 200,000 x g for 4 hours in a Sorvall M150 microcentrifuge. Equal volumes of fractions collected from the top of the gradient (20 µL) were separated by SDS-PAGE and analyzed by western blotting for the co-migration of CD63 and the indicated exosomal markers. (D) BT-CD63 cells (5×10<sup>6</sup>) were cultured in complete medium for 24 hours. Adherent cells were maintained in phenol red and exosome-free DMEM/F-12 for the indicated times (0–6 hours). For non-adherent cultures, EDTA-detached cells were resuspended in phenol red and exosome-free DMEM/F-12 for the indicated times (0–6 hours). GFP-CD63 tagged exosomes in the conditioned media were assayed by fluorescence spectroscopy. (E) Purified exosomes secreted for 2 hours from adherent or non-adherent BT-CD63 cells (5×10<sup>7</sup>) were separated in 4-12% SDS-polyacrylamide gels and analyzed by western blotting using antibodies against CD63 and the indicated exosomal markers.</p>", "links"=>[], "tags"=>["quantification", "intracellular", "released", "exosomes", "conditioned"], "article_id"=>408993, "categories"=>["Cancer", "Biochemistry", "Mathematics"], "users"=>["Rainelli B. Koumangoye", "Amos M. Sakwe", "J. Shawn Goodwin", "Tina Patel", "Josiah Ochieng"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0024234.g001", "stats"=>{"downloads"=>1, "page_views"=>23, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Detection_and_quantification_of_intracellular_and_released_exosomes_in_the_conditioned_medium_of_BT_CD63_/408993", "title"=>"Detection and quantification of intracellular and released exosomes in the conditioned medium of BT-CD63.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-09-06 02:29:53"}
  • {"files"=>["https://ndownloader.figshare.com/files/739076"], "description"=>"<p>The wells of a 96-well culture plate were either uncoated (plastic), or coated with 10 µg/ml of fibronectin or laminin for 16 hours at 37°C. Prior to use, the wells were washed twice with HBSS. BT-549 cells were pre-incubated in serum-free medium for 2 hours to deplete endogenous exosomes. The cells in serum free medium (SFM) were divided in 2 groups and re-suspended in SFM containing 1 mM Ca<sup>2+</sup>/Mg<sup>2+</sup> without or with cell-derived exosomes (10 µg/ml). Cells (2×10<sup>4</sup>) were transferred to 96-well plates for indicated times (30–120 minutes) and incubated at 37°C. The unattached cells were aspirated with the SFM and the adhered cells fixed in cold methanol and stained with crystal violet. Cells were photographed and the number of attached cells assessed by cell counting. Each bar represents the mean ± S.E of adherent cells per field. Bar is 40 µm.</p>", "links"=>[], "tags"=>["adhesion", "bt-549", "cancer"], "article_id"=>409449, "categories"=>["Cancer", "Biochemistry", "Mathematics"], "users"=>["Rainelli B. Koumangoye", "Amos M. Sakwe", "J. Shawn Goodwin", "Tina Patel", "Josiah Ochieng"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0024234.g004", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Exosomes_promotes_rapid_adhesion_of_BT_549_breast_cancer_cells_/409449", "title"=>"Exosomes promotes rapid adhesion of BT-549 breast cancer cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-09-06 02:37:29"}

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