Additional file 1 of GPR137-RAB8A activation promotes ovarian cancer development via the Hedgehog pathway

Supplementary Material 1: Figure S1. (A) The protein expression of GPR137 in SK-OV-3 cells transfected with RAB8A shRNA or scrambled shRNA (Con). (B) The protein expression of GPR137 in SK-OV-3 cells transfected with a RAB8A-expressing vector or a control vector (Con). Figure S2. (A) A heat-map of normalized expression levels of the altered genes measured by RNA-seq comparing an empty vector-transfected (Control) and a RAB8A-expressing vector-transfected (RAB8A) HO8910 cells. Blue indicates low expression levels and red indicates high expression levels. Figure S3. (A) The protein levels of GLI1 and PTCH1 in HO8910 cells transfected with a GPR137-expressing vector (GPR137, +) or a control vector (GPR137, -). Figure S4. (A) Matrigel invasion assays of SK-OV-3 (upper) and A2780 (lower) cells transfected with GPR137 shRNA (shGPR137) or scrambled shRNA (Control) in combination with SAG-treatment (SAG, +) or vehicle (SAG, -) for 24 h. Bar, 100 μm. (B) Quantitative analysis of (A, upper: SK-OV-3). (C) Quantitative analysis of (A, lower: A2780). (D) Colony formation assays of SK-OV-3 cells infected with lentiviruses carrying GPR137 shRNA or scrambled shRNA (shRNA-Con) in combination with SAG-treatment (SAG, +) or vehicle (SAG, -). (E) Colony formation assays of A2780 cells infected with lentiviruses carrying GPR137 shRNA or scrambled shRNA (shRNA-Con) in combination with SAG-treatment (SAG, +) or vehicle (SAG, -). (F) Quantitative analysis of relative colony numbers in (D). (G) Quantitative analysis of relative colony numbers in (E). (H) CCK-8 assays of HO8910 cells transfected with a GPR137-expressing vector or an empty vector (Control) in combination with GDC-0449-treatment (GDC) or vehicle and cultured for the indicated time periods. (I) Matrigel invasion assays of HO8910 cells transfected with a GPR137-expressing vector or an empty vector (Control) in combination with GDC-0449-treatment or vehicle for 24 h. Bar, 100 μm. (J) Quantitative analysis of (I). (K) Colony formation assays of HO8910 cells infected with lentiviruses expressing GPR137 or control lentiviruses in combination with GDC-0449-treatment. (L) Quantitative analysis of relative colony numbers in (K). (M) GLI luciferase (GLI-Luc) activities in HO8910 cells transfected with a GPR137-expressing vector or an empty vector (Control) in combination with RAB8A shRNA (shRAB8A, +) or scrambled shRNA (shRAB8A, -). (N) The mRNA levels of GLI1 and PTCH1 in HO8910 cells transfected with a GPR137-expressing vector or an empty vector (Control) in combination with RAB8A shRNA (shRAB8A, +) or scrambled shRNA (shRAB8A, -). (O) CCK-8 assays of HO8910 cells transfected with a GPR137-expressing vector or an empty vector (Control) in combination with RAB8A shRNA (shRAB8A) or scrambled shRNA and cultured for the indicated time periods. (P) Matrigel invasion assays of HO8910 cells transfected with a GPR137-expressing vector or an empty vector (Control) in combination with RAB8A shRNA (shRAB8A) or scrambled shRNA for 24 h. Bar, 100 μm. (Q) Quantitative analysis of (P). (R) Colony formation assays of HO8910 cells infected with lentiviruses expressing GPR137 or control lentiviruses in combination with lentiviruses expressing carrying RAB8A shRNA (shRAB8A) or scrambled shRNA. (S) Quantitative analysis of relative colony numbers in (R).