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. 2017 May 16;8:15237. doi: 10.1038/ncomms15237

Figure 6. The Src-dependent stress fibres and cell stiffening require EVL.

Figure 6

(a) Standard confocal sections of ER-Src cells treated with TAM or EtOH for 12 h and stained with β-CYA (magenta) and γ-CYA (green). (b) Standard confocal sections of ER-Src cells treated with TAM for 12 h and stained with Phalloidin (magenta), anti-EVL (green) and DAPI (blue). (c,d) Standard confocal sections of ER-Src cells treated with TAM or EtOH for 12 h, expressing shScr or shEVL#2, stained with Phalloidin (magenta), DAPI (blue) and (c) anti-Paxillin (yellow) or (d) anti-pMLC (yellow). All scale bars represent 10 μm. (e) Western blot on protein extracts from ER-Src cells treated with EtOH or TAM for 12 h, expressing shScr or shEVL#2, blotted with anti-actin to visualize the G- and F-actin pools. (f) Quantification of the G- (grey) and F-actin (magenta) ratio for the experimental conditions indicated. (g) Anisotropy of stress fibres in ER-Src cells expressing shScr (grey dots) or shEVL#2 (green dots) and treated with EtOH or TAM for 12 h. Each dot represents the anisotropy of single cells, with the horizontal line indicating median values. (h) Apparent Young's moduli of ER-Src cells expressing shScr (grey bars) or shEVL#2 (green bars) and treated with EtOH or TAM for 12 h. Data are presented as boxplots (25th, 50th, 75th percentiles), and whiskers indicate the 10th and 90th percentiles. All quantifications are from three biological replicates. Error bars indicate s.d.; NS indicates non-significant; *P<0.05; **P<0.001; ***P<0.0001. Statistical significance was calculated using (f,g) one-way ANOVA or (h) the Kruskal–Wallis test.