The m5C reader Ybx1 regulates embryonic cortical neurogenesis by promoting progenitor cell cycle progression
Fig 2
Conditional ablation of Ybx1 leads to a reduction in cortical progenitor cell pool and a decrease in cortical thickness.
(A) Immunostaining of coronal brain sections at P0 using antibodies against radial glial progenitor marker Pax6 and intermediate progenitor marker Tbr2. Representative cortical regions were shown. White dotted lines mark the boundaries. Scale bar, 100 μm. (B-F) Quantification of Pax6+ (B) and Tbr2+ (C) cell numbers, and cortical layer thicknesses (D–F) shown in (A). All statistical data are presented as box and whisker plots. For Pax6+ cell numbers, Ctrl (n = 26 confocal fields) vs. cKO (n = 24 confocal fields), ****p = 1.42E−05; Het (n = 23 confocal fields) vs. cKO, ****p = 9.54E−05. For Tbr2+ cell numbers, Ctrl (n = 26 confocal fields) vs. cKO (n = 24 confocal fields), ****p = 1.19E−05; Het (n = 23 confocal fields) vs. cKO, ***p = 1.85E−04. For VZ thickness, Ctrl (n = 26 confocal fields) vs. cKO (n = 24 confocal fields), ****p = 9.26E−05; Het (n = 23 confocal fields) vs. cKO, ****p = 1.33E−05. For SVZ thickness, Ctrl (n = 26 confocal fields) vs. cKO (n = 24 confocal fields), ***p = 9.49E−04; Het (n = 23 confocal fields) vs. cKO, **p = 0.0077. For whole cortical thickness, Ctrl (n = 26 confocal fields) vs. cKO (n = 24 confocal fields), ****p = 8.47E−06; Het (n = 23 confocal fields) vs. cKO, ****p = 2.61E−05. (G) Immunostaining of coronal brain sections at P0 for layer V marker Ctip2 and layer VI marker Tbr1. Representative cortical regions were shown. White dotted lines mark the boundaries of each layer. Scale bar, 100 μm. (H–M) Quantification of Ctip2+ (H) and Tbr1+ (I) neuron numbers, and the thickness of WM layer (J), layer VI (K), layer V (L), and layers I–IV (M) shown in (G). All statistical data are presented as box and whisker plots. For Ctip2+ neuron numbers, Ctrl (n = 22 confocal fields) vs. cKO (n = 20 confocal fields), ****p = 1.89E−06; Het (n = 20 confocal fields) vs. cKO, ***p = 1.62E−04. For Tbr1+ neuron numbers, Ctrl (n = 19 confocal fields) vs. cKO (n = 21 confocal fields), ****P = 4.62E−08; Het (n = 21 confocal fields) vs. cKO, ****P = 5.50E−05. For WM layer thickness, Ctrl (n = 22 confocal fields) vs. cKO (n = 20 confocal fields), ***p = 3.50E−04; Het (n = 20 confocal fields) vs. cKO, **p = 0.0038. For layer VI thickness, Ctrl (n = 22 confocal fields) vs. cKO (n = 20 confocal fields), ****p = 1.43E−05; Het (n = 20 confocal fields) vs. cKO, ***p = 2.56E−04. For layer V thickness, Ctrl (n = 22 confocal fields) vs. cKO (n = 20 confocal fields), ***p = 5.16E−04; Het (n = 20 confocal fields) vs. cKO, ***p = 5.71E−04. For thickness of layers, I–IV, Ctrl (n = 22 confocal fields) vs. cKO (n = 20 confocal fields), **p = 0.0093; Het (n = 20 confocal fields) vs. cKO, *p = 0.032. (N) A stacked bar chart summarizing the data in (D–F, J–M) shows the distribution of each layer in P0 cortex. At least 3 mice were analyzed for each genotype. All 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.