Figure 5

Specific second messenger blockade impacts on CPC characteristics. Application to CPCs of (a) Akt1 blocker A674563 or (b) p38 inhibitor SB203580 caused down-regulation of respective signalling pathways. (c) A674563 downregulated c-kit gene activity but did not impact on CPC ‘stemness’ genes or PDGFRα expression (n = 3), with (d) c-kit protein expression also assessed. (e) A674563 reduced expression of HGF mRNA (n = 3), with (f) protein expression also assessed. (g) SB203580 downregulated c-kit gene activity but did not impact on CPC ‘stemness’ genes or PDGFRα expression (n = 3), with (h) c-kit protein expression also assessed. (i) SB203580 significantly reduced EGF and HGF mRNA expression, with (j) protein expression also assessed. All data are mean ± SEM; significance was determined by ANOVA with Holm-Šídák post hoc test to identify differences (*p < 0.05; **p < 0.01; ***p < 0.001). Blots cropped to relevant data, with optical density normalized to the loading control protein; full blot images presented in Supplementary Fig. 6.