Fig. 2: FAK inhibition and SiRNA knockdown reduces CSC activity in MDA-MB-231 cells.

a Representative Western blot demonstrating that FAK inhibition with VS4718 resulted in a time dependent reduction in pTyr397FAK with relative protein expression shown in (b) but not FAK (n = 2). c Representative Western blot demonstrating pharmacological FAK inhibition with VS4718 resulted in a dose dependent reduction in pTyr397FAK with relative protein expression shown in (d) but not (e) FAK (n = 2). f The dose dependent reduction in pTyr397FAK corresponded with a reduction in primary MFE. g FAK knockdown using SiRNA resulted in a decrease in FAK expression with relative FAK: GAPDH after SiRNA knockdown shown in (h) relative density plot which correlated with a reduction in (i) primary MFE and (j) mammosphere self-renewal. One-way ANOVA with post hoc Dunnett’s test. k Representative FACS plots in scrambled controls and (l) SiRNA knockdown. m Graph demonstrating that FAK knockdown using SiRNA constructs 2 and 4 reduced ALDH⁺ expression (n = 2). Unpaired two-tailed t-test. (ns not significant, *p < 0.05, **p < 0.01, ***p < 0.001, ***p < 0.0001). All experiments were n = 3 unless otherwise stated and error bars represent mean + SEM.