Fig. 4: SEMA3G reduces c-Myc protein stability in GSCs.
From: Endothelial cells-derived SEMA3G suppresses glioblastoma stem cells by inducing c-Myc degradation
A Gene signatures enrichment of c-Myc targets in SEMA3G expressed GBM specimens. mRNA levels of c-Myc target genes in GSC07 (B) and GSC27 (C) cells treated with or without recombinant hSEMA3G (200 ng/ml) for 72 h. Data are shown as mean ± s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001. Data were analyzed by two-way ANOVA. The representative bands (D) and quantification (E) of c-Myc in GSC07 and GSC27 cells treated with or without recombinant hSEMA3G (200 ng/ml) for 72 h, as determined by western blot analysis. Data are shown as mean ± s.e.m. ***P < 0.001. Data were analyzed by two-tailed unpaired t test. F mRNA levels of c-Myc in GSC07 and GSC27 cells treated with or without recombinant hSEMA3G (200 ng/ml) for 72 h. Data are shown as mean ± s.e.m. Data were analyzed by two-tailed unpaired t test. G Correlation of SEMA3G and c-Myc at transcriptional level in GBM specimens from TCGA_GBMLGG dataset. The P values were calculated by the Pearson correlation test. H The representative bands of c-Myc in GSC07 and GSC27 cells treated with or without recombinant hSEMA3G (200 ng/ml) for 72 h, followed by CHX (10 µg/ml) for various time point. I, J Data summary of (H) are shown as mean ± s.e.m. **P < 0.01. Data were analyzed by two-way ANOVA. K GO terms analysis of the DEGs between GBM datasets (GSE162631) with different SEAM3G expression levels. L The representative bands of ubiquitination in GSC07 cells treated with SEMA3G (200 ng/ml) for 66 h, followed by MG132 (10 μmol/l) or the same volume DMSO for an additional 6 h. M The representative bands of the ubiquitination level of c-Myc in GSC07 cells treated with SEMA3G (200 ng/ml) for 72 h. The representative bands (N) and quantification (O, P) of c-Myc in GSC07 and GSC27 incubated with SEMA3G (200 ng/ml) for 66 h, followed by MG132 (10 μmol/l) for an additional 6 h. Data are shown as mean ± s.e.m. **P < 0.01. Data were analyzed by one-way ANOVA. All the western blot bands represent one of the three independent experiments.
