3.7 Generational Intersect of DMCs and DMRs and Top DMC Containing Genes
The overlap of DMC-containing genes was assessed as no single-nucleotide positional DMCs were shared between generations. Overall, few DMC-containing genes were shared between F1 and F2, with most overlapping genes occurring between iAs doses within-generation(Figure 8). The top three greatest DMC containing gene overlap was identified within generations and sex (Male F110 ppb vs 245 ppb, Female F2 10 ppb vs 245 ppb, and F1 Female 10 ppb vs. 245 ppb). We identified generational DMCs, finding males exposed to 245 ppb in utero had the highest overlap (8 DMCs) compared to other generational comparisons.
No DMR-containing genes were shared intergenerationally, although the genes Rlbp1 and Tcf4 both contained DMRs in 10ppb and 245ppb F1 males (Supplemental Table “DMR_all_hits.xlsx”). The Rlbp1 DMRs were hypomethylated and the Tcf4 DMRs were hypermethylated in both doses relative to control. One hypermethylated intronic DMC was identified in Tcf4 in F2 females at the 245ppb dose but not in any other F2 animals (Supplementary table “DML_all_hits.xlsx”. No Rlbp1 DMCs were observed in the F2 generation. This data shows limited persistence of DMCs and DMRs through the F1 to F2 generation.