Figure 5.
The anomaly of neurogenesis was rescued by overexpression of Mettl14 in primary NPCs from Arhgef2−/− mice (at E13.5)
(A) The protein expression of Npdc1 and Cend1 was examined by western blot. n = 3–4 per genotype, ∗p < 0.05, ∗∗p < 0.01, one-way ANOVA.
(B) The size of cortical neuro-spheres was measured at 72 h after transfection. A 2.3-fold increase of sphere size was noted by loss of Arhgef2, which could be recovered by HBLV-m-Mettl14-Zsgreen. Green: Zsgreen. Scale bar, 100 μm, n = 4 per genotype (3 measurements for each group), ∗∗p < 0.01, one-way ANOVA.
(C) Knockout of Arhgef2 significantly decreased βIII-tubulin+ cells (early neuron) and increased βIII-tubulin-Ki67+ proliferating cells (precursors), and the proportion could be rescued after Mettl14 overexpression. Cells were seeded at a density of 5 × 104/cm2 and were observed at day 4 after transfection. βIII-tubulin, marker for early neuron, green; Ki67, marker for proliferating cells, red; DAPI, blue. Scale bar, 50 μm. n = 6 per genotype, ∗∗p < 0.01, ∗∗∗p < 0.001, one-way ANOVA. Data are represented as mean ± SD.