Figure 4.

HLCs retain TET expression and 5hmC and demonstrate genic 5hmC enrichment in activated lipid synthesis and transport genes on LPO exposure. (a) 5hmC immune slot/blot of mouse liver, HLCs and HepG2 cells. Oligonucleotides of the APC gene promoter were used as controls. (b) qPCR of TET isoform mRNA expression during differentiation. Values are normalized to internal controls PPIA and B2M and expressed as fold change from undifferentiated ESCs. TET1 and TET3 expression decreases and TET2 expression increases during hepatic differentiation (*p < 0.05, **p < 0.01, ***p < 0.001 one-way ANOVA with Bonferroni multiple test correction versus ESCs). (c) Sliding window analysis of hmeDIP-seq displaying 5hmC profiles of HLCs stratified by expression quintile in relation to relative gene length (mean of two separate hmeDIP-seq experiments). (d) Sliding window analysis of all genes in control and LPO-treated HLCs (n = 3/group) shows no differences in global 5hmC levels. Error bars = s.d. (e) Heatmap analysis and unsupervised clustering of change in genic 5hmC specifically over induced genes of lipid synthesis and transport following LPO exposure.