Fig. 6: FAK inhibition reduces ex vivo mammosphere formation, ALDH⁺ expression and tumour initiating capacity.

a Graph demonstrating that FAK inhibition reduced ex vivo MFE when used alone or with Paclitaxel in RC37. b Graph demonstrating that VS4718 reduced MFE relative to control and Paclitaxel treated mice in RC193. Photomicrograph taken at ×40 magnification and 50 µM scale bars of mammospheres isolated ex vivo from; c RC37 control group, d RC37 VS4718 treated group, and e RC193 control receiving group. f Graph demonstrating a reduction in ALDH⁺ expression in VS4718 treated mice as opposed to Paclitaxel treated mice in RC37. g Graph demonstrating that VS4718 reduced ALDH⁺ in RC193 relative to control mice. Data in (a, b, f, and g) analysed using Two-way ANOVA with post hoc Dunnett’s test. h FACS plots demonstrating the identification of epithelial staining of ALDH was identified using PDX model. General cells were selected, then live cells isolated using 7AAD. Mouse cells were then excluded using H2KD. For the Aldefluor assay the DEAB control was set at 0.1%. With ALDH⁺ expression in control mice and VS4718 treated mice in the RC193 model shown in the last two panels. i Limiting dilution table demonstrating that combined FAK inhibition and Paclitaxel treatment led to a reduction in tumour initiating capacity. Tumours were defined based on size greater than 100 mm³ at Day 60 and ELDA analysis performed using online tool at: http://bioinf.wehi.edu.au/software/elda. j Combined FAK inhibition and Paclitaxel treatment resulted in a reduction in tumour size in the 100,000 cells per flank transplantation on Day 48. Data are represented as mean + SEM. (One-way ANOVA analysis with post hoc Dunnett’s test; *p < 0.05).