Extended Data Fig. 9: Menin and SUZ12 inhibition accelerates neurogenesis but not mouse neuroectoderm differentiation.
From: Genome-wide CRISPR screen identifies Menin and SUZ12 as regulators of human developmental timing
a, Schematic of dox-inducible KO at neuroectoderm stage. b, Representative Western Blot analysis to confirm the protein loss at the NPC (day 20) stage. c, Time course analysis of neuronal marker HuC/D and neural progenitor marker SOX2 with small molecule inhibitors (n = 3 independent experiments). d, Representative images of day 25 cells expressing neural progenitor marker SOX2. e, Quantification of MAP2 expression at day 25 of neurogenesis in small molecule inhibitors (left, n = 3 independent experiments), and NT and KOs (right, n = 3 independent experiments). P values were calculated using one-way ANOVA test with post-hoc Dunnett’s multiple comparisons test. f, Fold change of representative marker expression relative to control in EZH2i versus SUZ12 KO (n = 6 for EZH2i in DE; n = 3 for the rest). P values were calculated using two-tailed unpaired t-test. g, Representative images of H3K27me3 in EZH2i versus SUZ12 KO. h, Fold change of representative marker expression at differentiation endpoint with drug treatment (n = 2 for cTnT; n = 3 for ZBTB16, SOX2, HuCD; n = 4 for PAX6; n = 5 for GATA6, SOX17; n = 6 for CXCR4). i, Quantification of neural markers, Pax6 and Zbtb16, and a pluripotency marker, E-cadherin, during neuroectoderm differentiation of mouse epiblast stem cells (mEpiSCs), assessed by flow cytometry (n = 3 independent differentiations). No significant difference among means (P < 0.05) for all markers, tested by two-way ANOVA test with post-hoc Dunnett’s multiple comparisons test. j, Representative images of day 6 mouse cells expressing the neuroectoderm marker Pax6 and the neuronal marker Map2. White arrows point to Pax6-negative non-neural cells (top panels) and Map2-positive neurons (bottom panels). k, Venn diagrams show overlap in genes with H3K4me3 or MLL1 binding between hESCs and mEpiSCs. ChIP-seq data in mEpiSCs were downloaded from the ChIP-Atlas database93. All data are the mean ± s.d.
