Fig. 2

Zmynd11 deficiency disrupts neurogenesis in embryonic brain. A Representative WB images of Zmynd11, Tuj1 and Gfap in the brain of Zmynd11^loxp/loxp (Ctrl) and Nestin-Cre;Zmynd11^−/− (cKO) pups at the age of postnatal day 0 (P0). B, C Quantification results showed that cKO mice had decreased protein and mRNA levels of Zmynd11. n = 4 independent experiments for each group in (B) and n = 3 biologically independent experiments for each group in (C). Data represent mean ± SEM, unpaired t test; *, p < 0.05; **, p < 0.01; ***, p < 0.001; n.s. no significance. D, E Quantification results showed that cKO mice had decreased Gfap (D), but increased Tuj1 (E). n = 4 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. F Representative images of BrdU immunostaining with brain sections of Ctrl and cKO mice (P0). BrdU was injected (100 mg/kg, intraperitoneally [i.p.]) 1.5 h prior to the sacrifice. Scale bar, 50 μm. G Quantification results showed that the number of BrdU⁺ cells was significantly decreased in VZ/SVZ of cKO mice compared to Ctrl. n = 5 pups for each group. Data are presented as mean ± SEM; *p < 0.05; **p < 0.01; ***p < 0.001, ****p < 0.0001, unpaired Student’s t test. H Representative images of radial glial marker PAX6 immunostaining with brain sections of Ctrl and cKO mice at the age of P0. Scale bar, 50 μm. I Quantification results showed that the number of PAX6⁺ cells in VZ/SVZ was significantly decreased in VZ/SVZ of cKO mice compared to Ctrl. n = 5 pups for each group. Data are presented as mean ± SEM; *p < 0.05, **p < 0.01; ***p < 0.001, unpaired Student’s t test. J Representative images of intermediate progenitor cell marker Tbr2 immunostaining with brain sections of Ctrl and cKO mice (P0). Scale bar, 50 μm. K Quantification results showed the number of Tbr2⁺ cells was significantly increased in VZ/SVZ of cKO mice compared to Ctrl. n = 5 pups for each group. Data are presented as mean ± SEM; *p < 0.05, **p < 0.01, unpaired Student’s t test. L Representative images of BrdU immunostaining with scramble and Zmynd11 KD brain sections. Scramble and shZmynd11 plasmids were delivered into E14 fetal brain via in utero electroporation, and animals were sacrificed for assay at E17. BrdU was injected (100 mg/kg, i.p.) 1.5 h prior to the sacrifice. Scale bar, 50 μm. M Quantification results showed the percentage of BrdU⁺GFP⁺/GFP⁺ in VZ/SVZ was significantly decreased in Zmynd11 KD group compared to Ctrl. n = 5 fetuses at least from 4 pregnant mice for each group. Data are presented as mean ± SEM; *p < 0.05; **p < 0.01; ***p < 0.001, ****p < 0.0001, unpaired Student’s t test. N Representative images of Tuj1 immunostaining with scramble and Zmynd11 KD brain sections as in (L). Scale bar, 50 μm. O Quantification results showed the percentage of Tuj1⁺GFP⁺/GFP⁺ was significantly increased in cortical plate (CP) layer of Zmynd11 KD group compared to Ctrl. n = 5 fetuses at least from 4 pregnant mice for each group. Data are presented as mean ± SEM; *p < 0.05; **p < 0.01; ***p < 0.001, ****p < 0.0001, unpaired Student’s t test