Fig. 4

KPNB1 regulated glioblastoma progression through NLGN3. A. Western blot analysis showing KPNB1 and NLGN3 expression in U87MG and U251MG cells infected with lentivirus vectors expressing KPNB1-specific shRNAs. B. RT-qPCR results showing NLGN3 expression in U87MG and U251MG cells. Data presented as the mean ± SD of three independent experiments, ***P < 0.001, one-way ANOVA.C-D. U87MG and U251MG cells were infected with lentivirus vectors expressing KPNB1. C. Western blot analysis showing KPNB1 and NLGN3 expression. D. RT-qPCR results showing NLGN3 expression. Data presented as the mean ± SD of three independent experiments, ***P < 0.001, one-way ANOVA. E-L. U87MG and U251MG cells were infected with lentivirus vectors expressing shKPNB1, shNLGN3, or shKPNB1 combined with shNLGN3. Cells were collected for Western blot analysis (E), RT-qPCR (F), CCK8 assay (G), colony formation assay (H), and transwell invasion assay (I). J. After puromycin selection, cells were administered to nude mice by intracranial injection to establish the xenograft model. Representative bioluminescence imaging of a tumor at day 28 is shown. K. Quantitative assessments of tumor growth following implantation. Data expressed as mean + SD, n = 10 per group. *P < 0.05; **P < 0.01; ***P < 0.001, one-way ANOVA. L. Mice were sacrificed at ethical endpoint, survival curves were plotted in Kaplan–Meier graphs, and differences were evaluated using the log-rank test (P < 0.001 for control vs. shKPNB1; P < 0.001 for control vs. shNLGN3; P < 0.001 for control vs. shKPNB1 + shNLGN3; P = 0.057 for shKPNB1 vs. shKPNB1 + shNLGN3; P = 0.012 for shNLGN3 vs. shKPNB1 + shNLGN3)