Fig. 5

Non-mitochondrial p66ShcA accelerates the dynamics of focal adhesion formation. a Representative images of parental, p66-CR (VC), p66-CR (WT), and p66-CR (S36A)-expressing lung-metastatic 4T1 cells (537 population) transfected with mCherry paxillin. Scale bar is 10 μm. b Adhesions were segmented based on fluorescence intensity and analyzed for shape. Aspect ratio was determined by finding the ratio between semi-minor and semi-major axes. c Adhesions were segmented based on fluorescence intensity and analyzed for size. d Frequency distribution of adhesion areas from c. Data values were binned into 1-μm² segments. e Adhesions in protrusive cell regions were tracked over time to determine average assembly and disassembly rates from changes in mean fluorescence intensity. Cells were imaged every 20 s for a total of 25 min (parental: n = 12; VC: n = 11; WT: n = 10; S36A: n = 11). Data represent assembly and disassembly events for adhesions from three independent experiments. Top and bottom lines of the box indicate the 3rd and 1st quartile, respectively, while the bold central lines indicate mean. For panels b and c, statistical analysis was performed using a one-way ANOVA with a Tukey’s multiple comparisons test (****P < 0.0001). For panel e, statistical analysis was performed using a two-way ANOVA with a Tukey’s multiple comparisons test (****P < 0.0001). The whiskers extend up to 1.5 times the interquartile range