Figure 4

Relative transcript abundance of DNMT3A , DNMT3B , DNMT1 , TET family, UHRF1 family, TRIM28 / KAP1 and ZFP57 over five developmental stages. (A) The TPM values for each gene of interest were retrieved from a bovine embryo RNA-seq dataset (43) generated from embryos (n = 5) at Day 7 (blastocyst), Day 10 (post hatching spherical embryo/conceptus), Day 13 (ovoid embryo/conceptus), Day 16 (filamentous embryo/coneptus) and Day 19 (filamentous embryo/conceptus; initiation of implantation). Bars with similar superscripts (abcd) are not significantly different (P < 0.05). A significant effect of embryonic stage on gene expression was observed for DNMT3A, DNMT3B, DNMT1 and TRIM28/KAP1. Although a trend of decreasing transcript abundance is apparent for TET1-1, TET1-3 and ZFP57, the decrease was not statistically significant. Transcripts per million (TPM). (B) Western blot analysis of the DNA methyltransferases; DNMT3A, DNMT3A2 and DNMT3B. All proteins were expressed in Day 7 blastocyst samples (n = 4) with DNMT3A and DNMT3B being expressed in all Day 13 embryos (n = 4). DNMT3 methyltransferase protein expression was not detectable in Day 25 conceptuses (n = 4). A positive control (+Ctl) of total protein extracted from oocytes (shown previously to express the DNMT3 family members) was separated alongside the embryo samples.