Figure 4: Akt and RhoA pathways cooperate to activate NHE1 and stimulate migration in presence of cl-CD95L.

Cell migration of PS120-CD95 cells stably expressing wild type or ROCKA NHE1 mutants was assessed by wound healing assay in the presence or absence of cl-CD95 (100 ng/ml). Histogram bars correspond to the surface covered after 24 hours. Error bars are SEMs. p < 0.05 (*), p < 0.01 (**). (b) PS120-CD95 cells stably expressing the AKTD mutant were pre-treated with or without a non-cytotoxic dose of the ROCK1 inhibitor Y27632 (20 μM) for 1 h and then incubated for 24 h in the presence or absence of cl-CD95L (100 ng/ml). Migration was measured by wound healing assay. Histogram bars: surface area covered after 24 h. Error bars are the SEM. (c) Cell migration of PS120-CD95 cells stably expressing the AKTD-ROCKA NHE1 double mutants was measured by wound healing assay and compared to that of WT. Histogram bars correspond to the surface covered after 24 hours. Error bars are SEMs. p < 0.05 (*), p < 0.01 (**). (d) Summary of NHE1 activation by the non-conventional CD95 engagement.