Fig. 6.

ERK/p38 is downstream of RhoA/ROCK in supervillin-mediated cell metastasis during hypoxia. a. MHCC-97H cells that had been transfected with SV1, SV4, or SV5 plasmids were treated with PBS or the MEK inhibitor PD0325901 (10 μM) for 1 h under hypoxic conditions before assaying for phosphorylated p38, phosphorylated ERK, Snail1, and Vimentin by immunoblotting. β-tubulin was used as the loading control. b, c. MHCC-97H cells that had been transfected with SV1, SV4, or SV5 plasmids were treated with PBS or the MEK inhibitor PD0325901 (10 μM) for 1 h under hypoxic conditions before assay for cell migration (b) and invasion (c). The number of migrated SV1, SV4, or SV5 cells treated with PD0325901 was compared to those control cells treated with PBS. D. MHCC-97H and Huh-7 cells co-transfected with control or supervillin-specific siRNA and a RhoA(WT), RhoA(V14), or RhoA(N19) plasmids for 48 h were treated with PBS or the ROCK inhibitor Y27632 2HCl (10 μM) for 16 h during hypoxia, and then assayed for phosphorylated p38 and ERK by immunoblotting. β-actin was used as the loading control