Fig. 8: Depletion of PRC2 subunit Eed partially restores the appropriate gene expression and neuron differentiation in Ybx1-KO NPCs.
From: Ybx1 fine-tunes PRC2 activities to control embryonic brain development
a Genetic cross scheme to obtain embryos at E13.5 with Ybx1-KO or Ybx1-Eed-cKO by Nestin:Cre. WB analysis of Ybx1-KO NPCs with indicated Eed alleles (all embryos are Nestin:CreTg). b RT-qPCR with TaqMan assays of Ybx1-KO and Ybx1-Eed-cKO. Relative levels were displayed as their standardization to control NPCs. Dotted line indicates the level of gene expression in control cells. n = 6 except n ≥ 3 for Chrd, En2, and Shh. c IF analysis of cells with indicated genotypes at day 14 of differentiation. Quantification of d relative number of Tuj1-positive neurons, e neurite length, f number of branch points in axons, g number of non-trunk branches (not connected to an axon or dendrite), and h Gfap-positive (glial marker) cells in differentiating cells (n = 8 images). i RT-qPCR with TaqMan assays of neuronal genes in cells after 14 days of differentiation (n = 6). Dotted line indicates the level of gene expression in control cells. j Model of Ybx1 activities and its influence over PRC2 to ensure spatiotemporal precision in gene activation. Data are presented as mean ± SEM for b, d–i. P values by two-tailed unpaired t test are indicated for b and i and by ordinary one-way ANOVA followed by Tukey’s post hoc test in d–h. n.s indicates not significant. *, ***, and **** indicate P < 0.05, 0.001, and 0.0001, respectively. Source data are provided in Source Data file. P values: 8b(Fgf8)-0.0001, (Hes5)-0.0007, (Six3)-<0.0001, (Emx2)-<0.0001, (Arx)-<0.0001, (Bcl11b)-0.0006, (Fezf2)-<0.0001, (Dkk1)-0.003, (NeuroD2)-0.003, (Hoxa1)-<0.0001, (Msx1)-0.002, (Gbx2)-0.0002, (Chrd)-0.005, (Foxp2)-0.02, (En2)-<0.0001, (Foxa2)-<0.0001; 8d(Ybx1 WT-Eed cKO)-0.004, (Ybx1 WT-Ybx1 KO)-<0.0001, (Ybx1WT-dKO)-0.0003, (Eed cKO-Ybx1 KO)-0.0003, (Ybx1 KO-dKO)-0.004; 8e(Ybx1 WT-Ybx1 KO)-<0.0001, (Eed cKO-Ybx1 KO)-0.0003, (Ybx1 KO-dKO)-0.0002; 8f(Ybx1 WT-Ybx1 KO)-0.0042, (Eed cKO-Ybx1 KO)-0.008; ;8g(Eed cKO-Ybx1 KO)-0.0013; 8i(Satb2)-0.001, (NeuroD1)-0.0009, (Eno2)-<0.0001, (Tubb3)-0.01, (Map2)-<0.0001, (Dcx)-0.0002.
