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Fig. 2 | Breast Cancer Research

Fig. 2

From: Membrane associated collagen XIII promotes cancer metastasis and enhances anoikis resistance

Fig. 2

Collagen XIII promotes invasive 3D malignant phenotypes. a Schematic representation of human Col13A1 gene. Blue block represents exons. The area of the exon corresponding to the target region for CRISPR/Cas9 based gene editing is highlighted in red. The genomic sequencing results of the wildtype and mutant clones are presented in the box. A deletion was detected in Col13 knockout clone (Col13−/− (28)). b Western blot was performed to confirm Col13 knockout in Col13−/− (25) and Col13−/− (28) clones compared with control MDA-MB-231 cells. c Representative phase microscopy images of phenotype of MDA-MB-231 control and Col13−/− MDA-MB-231 cells under 3D matrix culture (left). Bar graph quantifying the invasive branches in MDA-MB-231 control and two Col13−/− clones (right). Results are presented as the mean ± s.e.m.; n = 100; *** p < 0.001. Scale bar: 50 μm. d Western blot analysis of flag-tagged Col13 and tubulin in cell lysate of control and Col13-expressing MCF-10A cells. e Fluorescence and phase images of control and col13-expression MCF-10A cells in 3D culture. Scale bars: 20 μm. f Line chart showing quantification of the diameter of control and Col13-expression MCF-10A cells at different time points in 3D culture. Data are presented as the mean ± s.e.m.; n = 60; * p < 0.05, *** p < 0.001; n.s., no significance. g Quantitative analysis of the cell proliferation of control and Col13-expression MCF-10A by EdU staining. Line chart representing the ratio of EdU positive to total cells. (day4, n = 20; day6, n = 20; day8, n = 20; day10, n = 20); *** p < 0.001; n.s., no significance

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