Fig. 3
c-Kit phosphorylates PHB in the membrane raft domain to enhance cancer cell migration. a Western blot analysis of c-Kit and PHB expression in the membrane raft or cytosolic plus non-raft membrane (C + M) fraction of SKOV3 and KURAMOCHI ovarian cancer cells. Caveolin-1 and clathrin heavy chain (HC) proteins are membrane raft and non-raft markers, respectively. b KURAMOCHI cells were treated with c-Kit inhibitor, imatinib mesylate (IM) for 24 h or siRNA for 72 h. Total cell lysates were subjected to western blotting. c SKOV3 cells transduced with lentiviral empty vector or vector encoding c-Kit were analyzed by western blotting of EMT markers. The migration ability was determined by 24-h Boyden chamber assay. Data represent the mean ± SD of three independent experiments. ***, P < 0.001, t test. d Analysis of c-Kit and PHB expression in the membrane raft or cytosolic plus non-raft membrane (C + M) fraction of SKOV3 and SKOV3_c-Kit cancer cells. e A SKOV3_c-Kit stable clone was transiently transfected with plasmid pD-PHBWT-HA that expresses exogenous PHB only in the membrane raft. The cell extracts were immunoprecipitated with anti-HA tag antibody, and coimmunoprecipitated proteins were detected by western blotting with anti-PHB and anti-c-Kit antibodies. Blots are representative of three independent experiments