Fig. 6: SEMA3G inactivates Cdc42 through NRP2/PLXNA1 in GSCs.

A Schematic illustration of the TurboID-tagged plasmid constructure and proximity labeling experiment. B PLXNA1, PLXNA2, PLXNA3, PLXNA4 and PLXND1 protein levels in whole cell lysates (input) or streptavidin bead–enriched protein samples from T98G cells transfected with NRP2-TurboID construct and treated with or without biotin. C HEK-293 cells were transfected with HA-tagged NRP2 and/or MYC-tagged PLXNA1. The interaction was determined using co-immunoprecipitation. D The PLXNA1 knockdown and control GSC07 and GSC27 cells treated with or without SEMA3G (200 ng/ml) for 72 h. The protein level of c-Myc was determined using western blot. E, F Data summary of (D) are shown as mean ± s.e.m. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001. Data were analyzed by one-way ANOVA. G PLXNA1 knockdown GSC07 cells were treated with or without SEMA3G (200 ng/ml) for 7 days. Images of neurospheres were captured using microscope. Scale bar, 100 μm. H Data summary of (G) are shown as mean ± s.e.m. ^***P < 0.001. Data were analyzed by one-way ANOVA. I The gene signatures of negative regulation GTPase enrichment analysis. J–O GSC07 and GSC27 cells were treated with or without SEMA3G (200 ng/ml) for 72 h. the activation of Cdc42 (J), RhoA (L), Rac1 (N) were determined using GST pulldown assay. K, M, O Data summary of (J, L, N) are shown as mean ± s.e.m. ^***P < 0.001. Data were analyzed by two-tailed paired t test. All the western blot bands represent one of the three independent experiments.