Skip to main content
. 2023 Aug 8;30(9):2053–2065. doi: 10.1038/s41418-023-01203-3

Fig. 6. Pd1 regulates Pax3 expression through AKT/GSK-3β/β-catenin signaling pathway.

Fig. 6

A Western blot analysis shows that Pd1 mediates the activation of the AKT/GSK-3β/β-catenin signaling pathway in brains. Proteins extracted from the brains of WT and cKO mice were probed with anti-p-AKT, anti-p-GSK3β(S9), anti-p-GSK3β(Y216), anti-p-β-catenin, anti-β-catenin, and anti-β-actin antibodies. B The level of β-catenin is increased when Pd1 is downregulated by shRNA. C The level of β-catenin is decreased obviously when both Pd1 and Pdl1 are overexpressed. D The level of β-catenin and Pax3 is reduced when XAV939 is added into the medium of NPCs. E The regulation of Pd1 on the AKT/GSK-3β/β-catenin pathway is mediated by SHP2. NPCs cultured in 6 well plates were infected with control and Pd1-sh1 lentivirus. Two days later, NPCs were treated with SHP099 at a concentration of 20 μM for 2 h at 37 °C and then harvested for the western blotting experiment. F Abnormal cell distribution caused by Pd1 suppression is rescued by the XAV939 administration. G Graph shows the percentage of GFP+ cells in each zone. n = 3 mice, p = 0.029 (DMSO-Control & DMSO-Pd1-sh1), p = 0.01 (DMSO-Control &XAV939-Control), p = 0.508 (XAV939-Control&XAV939-Pd1-sh1) for VZ/SVZ; p = 0.001 (DMSO-Control & DMSO-Pd1-sh1), p = 0.243 (DMSO-Control &XAV939-Control), p = 0.106 (XAV939-Control&XAV939-Pd1-sh1) for IZ; p = 0.002 (DMSO-Control & DMSO-Pd1-sh1), p = 0.005 (DMSO-Control &XAV939-Control), p = 0.732 (XAV939-Control&XAV939-Pd1-sh1) for CP. H Binding enrichment analysis of β-catenin on Pax3 promoter. The primary NSCs infected with lentivirus expressing β-catenin were subjected to chromatin immunoprecipitation experiments. n = 3 individual experiments, p = 0.59 for CDS; p = 0.436 for 500 bp; p = 0.024 for 1 kb; p = 0.713 for 2.5 kb. I Summary of Pd1 regulation on Pax3 via AKT/GSK-3β/β-catenin signaling pathway. Error bars represent means ± S.E.M.; Two-tailed unpaired t-test, P < 0.05(*), P < 0.01(**), n.s. not significant. The scale bar represents 100 μm.