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. 2020 Aug 14;219(9):e201912159. doi: 10.1083/jcb.201912159

Figure S2.

Figure S2.

CDC42EP5 promotes actomyosin function in collagen-rich matrices. (A) Images of 690.cl2 cells expressing GFP-Cdc42ep5 (green) on collagen-rich matrices. Merged and single greyscale channels also show F-actin (magenta) and pS19-MLC2 (cyan) staining. Scale bar, 50 µm. Violin plots show pS19-MLC2 (left) and F-actin (right) mean intensity from individual cells in 690.cl2 cells expressing different levels of Cdc42ep5 (high or low). n, individual cells; Mann–Whitney test. (B) Top panels represent the different shape of individual 690.cl2 cells and their respective roundness index. Left graph shows a violin plot of the roundness index of individual cells in 690.cl2 cells expressing different levels of Cdc42ep5 (high or low). n, individual cells; Mann–Whitney test. Right graph shows the relative frequencies of roundness indexes in Cdc42ep5hi and Cdc42ep5lo 690.cl2 cells. (C) Graph shows the relative frequencies of roundness indexes in 690.cl2 cells after transfection with control (siCtr) or two individual Cdc42ep5 (siEp5) siRNAs (additional representation of Fig. 2 C). (D) Graph shows the percentage of cells within three different ranges of roundness, from elongated (<0.4) to rounded (>0.6), in 690.cl2 cells after transfection with control (siCtr) or two individual Cdc42ep5 (siEp5) siRNAs. Bars represent mean. Additional representation of Figs. 2 C and S2 D. (E) Graphs show expression of CDC42EP1, CDC42EP2, CDC42EP3, and CDC42EP4 normalized to GAPDH expression in human melanoma cell lines with increasing rounding coefficients. Pearson’s correlation coefficient (r), statistical significance (p), and linear regression (red line) are shown. Each point in the graph represents the mean value of three independent experiments. (F) Western blot of indicated proteins in 690.cl2 cells after transfection with control (siCtr) and siRNAs targeting individual Borg genes (siEp1–5). Graph shows the quantification of normalized pS19-MLC2 levels in the different experimental points. Bars indicate mean ± SEM (n = 7 experiments; one-way paired ANOVA, Dunnett’s test: †, not significant; *, P < 0.05). AU, arbitrary units.