DNA Methylation may be a testicular plateau adaptation in Tibetan pig

Figures & data

Table 1. Primers used for real time PCR.

Gene names	GenBank accession	Primer pair sequences (5’ to 3’)	Product size (bp)
DNMT1	NM_001032355.1	F: ATGTGAGCCGAGCGAACT	110
		R: AGATGCCTGCTTGGTGGA
DNMT3A	NM_001097437	F: GATGGAAAGGGTGTTCGGCT	181
		R: TGTCACTACCTCGGTTTGCC
DNMT3B	XM_013985274.2	F: AGTAGGAGCGTAGAAGCAAG	244
		R: GGTGGAAAGACACGGGAT
HIF2α	NM_001097420	F: CCCACTGACAAGTTCATCCA	161
		R: GGGAGGCTGTAGTCCTGGTA
β-ACTIN	AJ312193.1	F:GCCAACAGAGAGAAGATGACAC	127
		R: GTAACACCATCACCAGAGTCCA

Table 2. Details of the antibodies used for western blots.

Name of antibody	Dilution ratios	Catalog #	Predicted band size	Producing areas
primary rabbit monoclonal DNMT1	1:1,000	ab13537	183 kDa	Abcam Inc., England
primary rabbit monoclonal DNMT3A	1:1,000	AF1732	120kDa	Beyotime Ltd., China
primary rabbit monoclonal DNMT3B	1:1,000	AF1384	96kDa	Beyotime Ltd., China
primary mouse monoclonal HIF2α	1:500	ab8365	100kDa	Abcam Inc., England
primary mouse monoclonal ACTIN	1:1,000	AA128	42kDa	Beyotime Ltd., China
HRP-labeled goat anti-mouse IgG(H + L)	1:1,000	A0216		Beyotime Ltd., China
HRP-labeled goat anti-rabbit IgG(H + L)	1:1,000	A0208		Beyotime Ltd., China

Figure 1. (a) The standard curve for quantifying genomic DNA methylation. (b) Genomic DNA methylation levels in testes. (c) Expression of DNA methyltransferases (DNMT1, DNMT3A, DNMT3B) and hypoxia-inducible factor 2α (HIF2α) mRNA in testes. (d) Western blot (WB) analysis of the four proteins (DNMT1, DNMT3A, DNMT3B and HIF2 α). ACTIN was used as a loading control. (e) Statistical results of WB for the four proteins in testes. Each bar represents mean ± standard error (S.E.), * Significant difference (P < 0.05), ** Extreme significant difference (P < 0.01). TP = Tibetan pig (n = 10); YP = Yorkshire pig (n = 10).