Fig. 3

DNA damage and activation of p53 and p53 target genes in Knl1 cKO brain. a Unique molecular identifier (UMI) RNA-seq volcano plot comparing cKO (n = 4) to ctrl (n = 7 animals) E15.5 cortex. P-value was calculated with likelihood ratio tests and false discovery rate (FDR) was calculated using the Benjamini–Hochberg procedure. Differentially expressed genes (FDR < 0.05) are indicated in red. b All 19 significantly upregulated genes. p53 target genes (pink), microglial genes (blue), and interneuronal genes (green) are indicated. c Droplet digital RT-PCR (ddRT-PCR) validated upregulation of p53 target genes Ano3, Eda2r, and Pvt1, and downregulation of Eomes in cKO compared to ctrl (mean, two-tailed unpaired t-test, ctrl: n = 7, cKO: n = 4 animals). d Immunostaining of E15.5 cortex revealed significant increases in the number of cells labeled by p53 or DNA damage marker γH2AX in cKO compared to ctrl (mean, two-tailed unpaired t-test, n = 3 animals, scale bar: 100 µm). p53 was localized to nuclei (solid arrowheads) and present in pyknotic cells (open arrowheads, inset scale bar: 5 µm). e Double immunostaining revealed p53 activation in SOX2+ NPCs in E15.5 cKO VZ (arrowheads). The vast majority of cells (89 ± 1.5%) with p53 activation were SOX2+ NPCs (mean, cKO: n = 4 animals). coloc. colocalization