Fig. 6

Exogenous Epha2 rescues neurogenic deficits and restores PI3K signaling in Zmynd11 KD eNPCs. A Representative images of BrdU immunostaining of WT eNPCs infected with lenti-scramble-GFP, lenti-shZmynd11-GFP, and lenti-shZmynd11-GFP + lenti-Epha2-RFP, respectively. BrdU was incorporated for 6 h at a final concentration of 5 μM. Scale bar, 50 μm. B Quantification results showed that ectopic Epha2 significantly enhanced the percentage of BrdU⁺ cells in Zmynd11 KD eNPCs compared to Ctrl. n = 4 biologically independent experiments. Data are presented as mean ± SEM; *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001, one-way ANOVA analysis followed by Tukey’s multiple-comparison test, F_{(2, 9)} = 13.73. C Representative images of Tuj1 immunostaining of WT eNPCs infected with lenti-scramble-GFP, lenti-shZmynd11-GFP, and lenti-shZmynd11-GFP + lenti-Epha2-RFP, respectively. Scale bar, 50 μm. D Quantification results showed that ectopic Epha2 significantly reduced the percentage of Tuj1⁺ cells in Zmynd11 KD group compared to Ctrl. n = 4 biologically independent experiments. Data are presented as mean ± SEM; *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001, one-way ANOVA analysis followed by Tukey’s multiple-comparison test, F_{(2, 9)} = 154.6. E Representative images of immunostaining of WT eNPCs infected with lenti-scramble-GFP, lenti-shZmynd11-GFP, and lenti-shZmynd11-GFP + lenti-Epha2-RFP, respectively. Scale bar, 50 μm. F Quantification results showed that ectopic Epha2 significantly enhanced the percentage of Gfap⁺ cells in Zmynd11 KD group compared to Ctrl. n = 4 biologically independent experiments. Data are presented as mean ± SEM; *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001, one-way ANOVA analysis followed by Tukey’s multiple-comparison test, F_{(2, 9)} = 59.39. G–M Representative WB images (G) and quantification results showed that exogenous Epha2 did not affect the protein level of Zmynd11 (H), but significantly enhanced the level of Epha2 (I) in Zmynd11 KD eNPCs compared to Ctrl. Exogenous Epha2 significantly enhanced p-PI3K/PI3K (J), p-SGK1/SGK1 (K), and Gfap (L), but reduced the level of Tuj1 (M) in Zmynd11 KD eNPCs compared to Ctrl. n = 4 biologically independent experiments. Data are presented as mean ± SEM; *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001, one-way ANOVA analysis followed by Tukey’s multiple-comparison test, F_{(2, 12)} = 30.36 for (H), F _{(2, 12)} = 61.28 for (I), F _{(2, 12)} = 12.68 for (J), F _{(2, 12)} = 50.39 for (K), F _{(2, 12)} = 40.36 for (L), F _{(2, 12)} = 15.66 for (M)