Fig. 8

GOLM1 may mediate PDGFA/PDGFRα signaling in A172 cells in vitro. a Western blot analysis of p-PDGFRα in lysates prepared from PDGFA treated (20 ng/mL) and untreated U251 and A172 cells. PBS was used as the vehicle control. b IF staining of GOLM1 (red) in A172 cells treated with 0 ng/mL, 20 ng/mL and 50 ng/mL PDGFA for 24 h. Nuclei were labeled with DAPI (blue). Scale bar = 20 μm. c qRT-PCR (upper) and Western blot analysis (lower) of GOLM1 in lysates prepared from A172 cells treated with 0, 20, and 50 ng/mL PDGFA for 48 h. d qRT-PCR (upper) and Western blot analysis (lower) of GOLM1 in untreated cells or cells treated with increasing amounts of PDGFA in the presence of DMSO (vehicle control) or an inhibitor of PDGFRα AG1296 (5 μM) for 48 h. A172-NC and -sh-GOLM1 cells were treated with PBS (0 ng/mL PDGFA) as negative control or PDGFA (20 ng/mL) for 48 h. e EdU assays to evaluate cell proliferation under indicated treatment. Scale bar = 100 μm. f Representative images of Transwell migration and invasion assays performed in cells with indicated treatment. g Graphic representation of ratios of EdU positive cells under different treatments. Data are presented as the mean ± SEM. h Quantification of invaded and migrated cells in Transwell assays after incubation for 24 h. Data is presented as the mean ± SEM. Scale bar = 50 μm. i Western blot analysis of p-AKT (S473), AKT, p-ERK1/2 and ERK in A172-NC and -sh-GOLM1 cells after treatment with PDGFA (20 μg/mL) for 0, 10, 20, and 30 min. (NS, not significant; *P < 0.05, **P < 0.01, ***P < 0.001)