Fig. 6
Involvement of YAP1 in LNCaP cell migration. A Migration of YAP1 knockout LNCaP cells. Migratory cell number was determined by IncuCyte ZOOM live-cell analysis system. Data represent means ± SD (n = 4, *P < 0.05 vs control cell). B YAP/TAZ-reporter activity of YAP knockout LNCaP cells. Control and YAP knockout LNCaP cells were transiently transfected with YAP/TAZ-reporter plasmid (TEAD reporter, 1 μg/ml) and the cells were incubated with or without 10% FBS for 24 h. Luciferase activity was determined to assess YAP/TAZ-mediated gene transcription. Data represent means ± SD (n = 6, **P < 0.01 vs control LNCaP cells). C Diminished cell migration by YAP knockout in serum-containing and serum-deprived conditions. Data represent means ± SD (n = 4, *P < 0.05, **P < 0.01 vs vehicle-treated control). D Effect of GV1001 on YAP1 knockout LNCaP cell migration. Control- and YAP1-knockout LNCaP cells were treated with 10 μM GV1001 for 24 h, and cell migration was quantified by IncuCyte ZOOM live-cell analysis system. Data represent means ± SD (n = 4, *P < 0.05 vs vehicle-treated control). E Establishment of constitutive active YAP5SA-LNCaP cells. Protein expression of YAP1, Myc, and CTGF were determined by immunoblot analyses. F Effect of GV1001 on the migration of YAP5SA-overexpressing LNCaP cells. Control and YAP-5SA cells were exposed to 10 μM GV1001 for 24 h. Data represent means ± SD (n = 4, **P < 0.01 vs vehicle-treated control, ##P < 0.01 vs control-LNCaP cell group). G Forskolin-induced YAP1 phosphorylation. LNCaP cells were treated with 0.1–10 μM Forskolin for 1 h, and protein levels of YAP1 and p-YAP1 were determined by immunoblot analysis. H Effect of forskolin on LNCaP cell migration. LNCaP cells were treated with forskolin (10 μM) for 24 h. Data represent means ± SD (n = 4, **P < 0.01 vs vehicle-treated control). I Reversal of GV1001-induced anti-migratory activity by KT5720 and KH7. LNCaP cells were incubated with 10 μM GV1001 in the presence or absence of KT5720 (1 μM) or KH7 (1 μM) for 24 h. Data represent means ± SD (n = 4, *P < 0.05, **P < 0.01 vs vehicle-treated control, ###P < 0.005 vs GV1001-treated group)