The m5C reader Ybx1 regulates embryonic cortical neurogenesis by promoting progenitor cell cycle progression
Fig 3
Ybx1 is required for maintenance of the cortical progenitor pool from E13.5.
(A) E13.5 coronal brain sections were immunostained with antibodies against the radial glial cell marker Pax6 and the intermediate progenitor marker Tbr2. Representative cortical regions were shown. White dotted lines mark the boundaries. Scale bar: 50 μm. (B–F) Quantification of Pax6+ (B) and Tbr2+ (C) cell numbers, and cortical layer thicknesses (D to F) shown in (A). All statistical data are presented as box and whisker plots. For Pax6, Ctrl (n = 21 confocal fields) vs. cKO (n = 22 confocal fields), **p = 0.0018; Het (n = 18 confocal fields) vs. cKO, *p = 0.019. For Tbr2, Ctrl (n = 21 confocal fields) vs. cKO (n = 22 confocal fields), *p = 0.025; Het (n = 18 confocal fields) vs. cKO, *p = 0.014. For whole cortical thickness, Ctrl (n = 21 confocal fields) vs. cKO (n = 22 confocal fields), **p = 0.0018; Het (n = 18 confocal fields) vs. cKO, *p = 0.016. For VZ thickness, Ctrl (n = 21 confocal fields) vs. cKO (n = 22 confocal fields), **p = 0.0023; Het (n = 18 confocal fields) vs. cKO, *p = 0.026. For SVZ thickness, Ctrl (n = 21 confocal fields) vs. cKO (n = 22 confocal fields), **p = 0.0055; Het (n = 18 confocal fields) vs. cKO, *p = 0.047. (G) A stacked bar chart summarizing the data in (D–F) shows the distribution of each layer in E13.5 cortex. (H) E15.5 coronal brain sections were immunostained with antibodies against the radial glial cell marker Pax6 and the intermediate progenitor marker Tbr2. Representative cortical regions were shown. White dotted lines mark the boundaries. Scale bar: 100 μm. (I–O) Quantification of Pax6+ (I) and Tbr2+ (J) cell numbers, and cortical layer thicknesses (K–O) shown in (H). All statistical data are presented as box and whisker plots. For Pax6, Ctrl (n = 24 confocal fields) vs. cKO (n = 22 confocal fields), ***p = 2.93E−04; Het (n = 21 confocal fields) vs. cKO, **p = 0.0071. For Tbr2, Ctrl (n = 24 confocal fields) vs. cKO (n = 22 confocal fields), ***p = 8.58E−04; Het (n = 21 confocal fields) vs. cKO, **p = 0.0042. For VZ thickness, Ctrl (n = 24 confocal fields) vs. cKO (n = 22 confocal fields), ****p = 5.60E−05; Het (n = 21 confocal fields) vs. cKO, ***p = 2.72E−04. For SVZ thickness, Ctrl (n = 24 confocal fields) vs. cKO (n = 22 confocal fields), ***p = 2.75E−04; Het (n = 21 confocal fields) vs. cKO, ***p = 8.94E−04. For CP thickness, Ctrl (n = 24 confocal fields) vs. cKO (n = 22 confocal fields), **p = 0.0062; Het (n = 21 confocal fields) vs. cKO, *p = 0.034. For IZ thickness, Ctrl (n = 24 confocal fields) vs. cKO (n = 22 confocal fields), ***p = 1.99E−04; Het (n = 24 confocal fields) vs. cKO, ***p = 5.31E−04. For whole cortical thickness, Ctrl (n = 24 confocal fields) vs. cKO (n = 22 confocal fields), ****p = 1.73E−05; Het (n = 21 confocal fields) vs. cKO, ***p = 1.68E−04. (P) A stacked bar chart summarizing the data in (K–O) shows the distribution of each layer in E15.5 cortex. At least 3 embryos 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.