Figure 8

SOX2-RNAi-induced membrane protrusions are associated with increased RhoA activity and are repressed by blocking ROCK. a: RhoA-G-Lisa of U343-MG and U373-MG cells with knockdown of SOX2 compared to shLuc control cells. Protein lysates were prepared and subjected to RhoA-G-LISA. Inlays: RhoA-input in G-LISA. *p < 0.05 when cells with knockdown of SOX2 were compared to the shLuc controls. b: Inhibition of ROCK signaling downstream of RhoA with the small chemical compound Y27632 completely abolish membrane blebbing in U343-MG with knockdown of SOX2. The inhibitor was added at time 00.00.00 (hh:mm:ss). c: Western blot analysis confirms ROCK2 expression in U343-MG and U373-MG cells with knock down of SOX2 which is not affected by treatment with Y27632. The inhibition of ROCK led to diminished levels of phosphorylated myosin light chain (MLC-P). Equal loading of the filters was confirmed by stripping the filters and probing with tubulin antibodies. Densitometric analyses of Western blot signals normalized to tubulin signals are depicted.