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. 2020 May 15;11:2416. doi: 10.1038/s41467-020-16199-4

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a Doxorubicin response of MDA-MB-231 cells cultured with or without type I collagen for 72 h (n = 4). b Apoptosis assay by Annexin V/DAPI staining from a (n = 2). c Western blot analysis in doxorubicin-treated cells grown with or without type I collagen. d Percentage growth inhibition in LOX-overexpressing cells embedded in collagen upon doxorubicin treatment (n = 3). e Western blot analyses upon LOX overexpression in MDA-MB-231 cells. f, g Percentage growth inhibition of collagen-embedded MDA-MB-231 cells treated with BAPN (f) or transfected with siLOX (g) in combination with doxorubicin (n = 3). h, i Immunofluorescence staining (h) and quantifications of the intensities (i) of extracellular type I collagen and fibronectin upon treatment of collagen-embedded cells with BAPN (n = 18 (vehicle), n = 15 (BAPN) for collagen, and n = 18 (vehicle), n = 12 (BAPN) for fibronectin). j, k Immunofluorescence staining (j) and quantifications of the intensities (k) of HFF-derived type I collagen and fibronectin incubated with vehicle or BAPN-treated MDA-MB-231 cells. ECM without cells represents the staining in the absence of MDA-MB-231 cells (n = 29 (vehicle), n = 25 (BAPN)). l Amount of cross-linked collagen in HFF-derived ECM incubated with vehicle- vs. BAPN-treated MDA-MB-231 cells (n = 3 different wells). m Western blot of soluble and insoluble FN1 obtained by deoxycholate lysis of NIH3T3- and HFF-derived ECM in contact with vehicle- vs. BAPN-treated MDA-MB-231 cells. n Changes in relative doxorubicin fluorescence upon BAPN-treatment in MDA-MB-231 cells embedded in type I collagen (n = 5). o Western blot analysis of LOX and FAK/Src signaling in collagen type I-embedded MDA-MB-231 cells upon doxorubicin and BAPN treatment for 24 h. p Annexin V/DAPI staining upon combination treatment for 72 h (n = 2). q Percentage growth inhibition induced by the combination of doxorubicin with FAK (PF-562271) or Src (Saracatinib) inhibitors in MDA-MB-231 cells embedded in type I collagen (n = 3). Data represents mean ± SD. Two-sided Student’s t-test was used to calculate statistical difference between two groups. One-way ANOVA with Dunnett’s test was performed to compare mean of combination-treated group with single agent treatments in f, q. Scale bar = 50 µm for h, j. Source data are provided as a Source data file.