5mC-loss/5hmC-gain loci characterize neurogenic genes up-regulated during fate commitment. 5mC-loss/5hmC-gain loci characterize neurogenic genes up-regulated.

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5mC-loss/5hmC-gain loci characterize neurogenic genes up-regulated during fate commitment. 5mC-loss/5hmC-gain loci characterize neurogenic genes up-regulated during fate commitment. (A) Volcano plots depicting the loss (orange) and gain (blue) in 5mC (left) or 5hmC (right) from PP to DP and DP to neurons (as indicated). Number of significantly (>50% change, P < 0.05) differentially (hydroxy-)methylated loci (top) and nearest genes (parentheses) are indicated. (B, C) Heat maps (left) and violin plots (right) representing 5(h)mC levels within a ±2-kb region across differentially (hydroxy-)methylated loci in each cell type (colors, as above) calculated as log2(ChIP/control) or trimmed mean of M values (TMM), respectively. In this analysis, 5hmC-gain and 5mC-loss loci identified in DP (4,757) or neurons (3,716), respectively, were first taken as a reference (top panels) and the levels of the converse cytosine modification (i.e., 5mC or 5hmC for B and C, respectively) measured within these very same loci (bottom) revealing the extensive correlation in 5(h)mC changes. (D) Whiskers–box plots of 5mC and 5hmC levels (left and right, respectively) calculated for individual cytosines (n = 48) after (oxidative) bisulfite amplicon sequencing of six among all 5mC-loss/5hmC-gain loci (one examples of which is depicted in Fig S2C). Data are plotted as fold change relative to PP. (E, F) DAVID-gene ontology term enrichment (top three terms) (E) and expression levels (FPKM) (F) of 5mC-loss and 5hmC-gain loci in the PP–DP and DP–neuron transitions (as indicated). Note the high enrichment, specificity, and consistency of the GO terms and the highly significant change in gene expression throughout the neurogenic lineage. Statistical test = edgeR-modified t test (A), Wilcoxon rank sum test (B, C, D and F). Florian Noack et al. LSA 2019;2:e201900331 © 2019 Noack et al.