The m5C reader Ybx1 regulates embryonic cortical neurogenesis by promoting progenitor cell cycle progression
Fig 7
Overexpression of Ybx1 targets rescues the cortical development defects caused by Ybx1 ablation.
(A) Overexpression of Ybx1 targets (OE-Cocktail) in Ybx1 cKO mouse cortex using IUE, followed by immunostaining for GFP, Pax6, and Tbr2 on coronal sections at E16.5. Representative cortical regions were shown. Scale bar, 50 μm. (B and C) Quantification of percentage of Pax6+GFP+/GFP+ (B) and Tbr2+GFP+/GFP+ (C) shown in (A). Data are presented as box and whisker plots. In B, “Ctrl + OE-Ctrl” (n = 20 confocal fields) vs. “cKO + OE-Ctrl” (n = 18 confocal fields), ****p = 4.26E−05; “cKO + OE-Ctrl” vs. “cKO + OE-Cocktail” (n = 21 confocal fields), **p = 0.0040; “Ctrl + OE-Ctrl” vs. “cKO + OE-Cocktail”, p = 0.10. In C, “Ctrl + OE-Ctrl” (n = 20 confocal fields) vs. “cKO + OE-Ctrl” (n = 18 confocal fields), ***p = 5.69E-04; “cKO + OE-Ctrl” vs. “cKO + OE-Cocktail” (n = 21 confocal fields), *p = 0.043; “Ctrl + OE-Ctrl” vs. “cKO + OE-Cocktail”, *p = 0.015. (D) Overexpression of Ybx1 targets (OE-Cocktail) in control and Ybx1 cKO mouse cortex using IUE, followed by immunostaining for GFP, EdU, and Ki67 on coronal sections at E16.5. Representative cortical regions were shown. Scale bar, 50 μm. (E and F) Quantification of percentage of EdU+GFP+/GFP+ (E) and Ki67-EdU+/EdU+GFP+ (F) shown in (D). Data are presented as box and whisker plots. In E, “Ctrl + OE-Ctrl” (n = 27 confocal fields) vs. “Ctrl + OE-Cocktail” (n = 25 confocal fields), **p = 0.0099; “Ctrl + OE-Ctrl” vs. “cKO + OE-Ctrl” (n = 24 confocal fields), ****p = 3.08E−10; “Ctrl + OE-Ctrl” vs. “cKO + OE-Cocktail” (n = 26 confocal fields), p = 0.19. “Ctrl + OE-Cocktail” vs. “cKO + OE-Cocktail”, ****p = 8.25E−6. “cKO + OE-Ctrl” vs. “cKO + OE-Cocktail”, ****p = 3.08E−10. In F, “Ctrl + OE-Ctrl” (n = 26 confocal fields) vs. “Ctrl + OE-Cocktail” (n = 28 confocal fields), *p = 0.015; “Ctrl + OE-Ctrl” (n = 26 confocal fields) vs. “cKO + OE-Ctrl” (n = 24 confocal fields), ****p = 3.08E−10; “Ctrl + OE-Ctrl” vs. “cKO + OE-Cocktail” (n = 24 confocal fields), p = 0.51. “Ctrl + OE-Cocktail” vs. “cKO + OE-Cocktail”, ***p = 0.00015. “cKO + OE-Ctrl” vs. “cKO +OE-Cocktail”, ****p = 3.08E−10. At least 3 embryos were analyzed for each genotype or condition. Analyses were performed by one-way ANOVA followed by Tukey’s multiple comparison test. ns, not significant. The data underlying all the graphs shown in the figure are included in S1 Data.