Fig. 4

Identification of HH signaling as the downstream of RAB8A. A The protein levels of exogenous RAB8A (Flag) in RAB8A cells transfected with a Flag-tagged RAB8A (Flag-RAB8A, +) or an empty vector (Flag-RAB8A, -) for 24 h were detected by western blot. B The number of genes whose expression was altered by Flag-RAB8A-expression is summarized in the table. C Volcano scatter plot for the genes in (B). D GO analysis was conducted to compare the gene expression from RAB8A group and the control. E KEGG pathway enrichment analysis was conducted to compare the gene expression from RAB8A group and the control. F The protein expression of GLI1 and PTCH1 in HO8910 cells transfected with RAB8A (Flag-RAB8A, +) or an empty vector (Flag-RAB8A, -). G GLI luciferase (GLI-Luc) activities in HO8910 cells transfected with RAB8A or an empty vector (Con). H Nude mice with xenografts derived from HO8910 cells stably expressing RAB8A or control viruses. I Image showing the size of the representative tumor xenografts from two groups. J Weights of xenografts derived from HO8910 cells stably expressing RAB8A or control viruses at 21 d post inoculation. K The protein levels of GLI1 and RAB8A in tumors from HO8910 cells stably expressing RAB8A or control viruses. L The protein expression of PTCH1, GLI1, GLI2, GLI3-full length (GLI3F), GLI3-repressor (GLI3R), IFT88, KIF3A, and SuFu in SK-OV-3 cells transfected with RAB8A shRNA (shRAB8A, +) or scrambled shRNA (shRAB8A, -). M The protein expression of GLI1 and PTCH1 in A2780 cells transfected with RAB8A shRNA (shRAB8A, +) or scrambled shRNA (shRAB8A, -). N GLI luciferase (GLI-Luc) activities in SK-OV-3 cells transfected with RAB8A shRNA (shRNA RAB8A) or scrambled shRNA (shRNA Con). O GLI luciferase (GLI-Luc) activities in A2780 cells transfected with RAB8A shRNA (shRNA RAB8A) or scrambled shRNA (shRNA Con). P The mRNA levels of GLI1 and PTCH1 in SK-OV-3 cells transfected with RAB8A shRNA (shRNA-RAB8A) or scrambled shRNA (shRNA-Con). Q The mRNA levels of GLI1 and PTCH1 in A2780 cells transfected with RAB8A shRNA (shRNA-RAB8A) or scrambled shRNA (shRNA-Con). R Co-immunoprecipitation of endogenous SuFu and GLI1 in the presence of RAB8A shRNA (shRAB8A) in SK-OV-3 cells. IP: SuFu; WB: GLI1. IgG was used as a negative control. S Co-immunoprecipitation of endogenous SuFu and GLI1 in the presence of ectopic Flag-RAB8A in HO8910 cells. IP: SuFu; WB: GLI1. IgG was used as a negative control. T Nucleo-cytoplasmic separation assays in HO8910 cells transfected with Flag-RAB8A or an empty vector (Flag-RAB8A, -), and protein levels of GLI1 were detected by western blot. U GEPIA database displayed the correlation between RAB8A and PTCH1. *p < 0.05; **p < 0.01; error bar, SD