Fig. 1

Zmynd11 depletion impairs the proliferation and differentiation of eNPCs in vitro. A–D Representative Western blot (WB) images of Zmynd11, neuronal marker Tuj1 and astrocytic marker Gfap (A). Zmynd11 KD significantly reduced the level Zmynd11 protein in proliferating eNPCs (B). Under differentiation condition, Zmynd11 KD significantly reduced Gfap (C), but increased Tuj1 protein compared to scramble group (Ctrl) (D). n = 3 biologically independent experiments for each group. Data represent mean ± SEM, unpaired t test; *, p < 0.05; **, p < 0.01; ***, p < 0.001; n.s. no significance. E Representative images of Ki67 immunofluorescence staining with eNPCs infected with lentivirus expressing scramble and shRNA against Zmynd11 (shZmynd11) compared to scramble group (Ctrl). eNPCs were infected with lentivirus for 72 h. Scale bar, 50 μm. F Quantification results showed that Zmynd11 knockdown (KD) led to a decreased percentage of Ki67⁺ cells. n = 4 biologically independent experiments for each group. Data represent mean ± SEM, unpaired t test; *, p < 0.05; **, p < 0.01; ***, p < 0.001; n.s. no significance. G Representative images of BrdU immunofluorescence staining with eNPCs infected with lentivirus expressing scramble and shZmynd11. eNPCs were infected with lentivirus for 72 h, and BrdU was incorporated for 6 h at a final concentration of 5 μM. Scale bar, 50 μm. H Quantification results showed that Zmynd11 KD reduced the percentage of BrdU⁺ cells. n = 4 biologically independent experiments for each group. Data represent mean ± SEM, unpaired t test; *, p < 0.05; **, p < 0.01; ***, p < 0.001; n.s. no significance. I Representative images of Tuj1 immunofluorescence staining with eNPCs infected with lenti-scramble and lenti-shZmynd11. After eNPCs were infected with lentivirus for 72 h, eNPCs differentiation was induced for 48 h. Scale bar, 50 μm. J Quantification results showed that Zmynd11 KD enhanced the percentage of Tuj1⁺ cells. n = 4 biologically independent experiments for each group. Data represent mean ± SEM, unpaired t test; *, p < 0.05; **, p < 0.01; ***, p < 0.001; n.s. no significance. K Representative images of Gfap immunofluorescence staining with eNPCs infected with lenti-scramble and lenti-shZmynd11. After eNPCs were infected with lentivirus for 72 h, eNPCs differentiation was induced for 48 h. Scale bar, 50 μm. L Quantification results showed that Zmynd11 KD reduced the percentage of Gfap⁺ cells. n = 4 biologically independent experiments for each group. Data represent mean ± SEM, unpaired t test; *, p < 0.05; **, p < 0.01; ***, p < 0.001; n.s. no significance. M–P qRT-PCR assay results showed that Zmynd11 KD significantly reduced the mRNA levels of Zmynd11 (M), Gfap (N), s100β (O) and increased the mRNA level of Tuj1 (P). n = 3 independent experiments for each group. Data represent mean ± SEM, unpaired t test; *, p < 0.05; **, p < 0.01; ***, p < 0.001; n.s. no significance