Fig. 5

PDE induced the region hypermethylation change of the LOX promoter via GR and ERβ interactions. A-F Bisulfite Sequencing PCR of methylation levels in the LOX promoter region in bone tissue of fetal rats and adult offspring rats and osteoclasts treated with 500 nM DEX. G, H Screening of methyltransferases in bone tissue of fetal rats and osteoclasts treated with 500 nM DEX. I Tet3 protein expression in osteoclasts treated with 500 nM DEX. J Quantification analyses of Tet3 protein expression. K, L CHIP-PCR for the targeted binding of GR to LOX in bone tissue of fetal rats and osteoclasts treated with 500 nM DEX. M, N GR and ERβ mRNA expression in bone tissue of fetal rats and osteoclasts treated with 500 nM DEX. O Co-IP for GR and ERβ interactions in osteoclasts treated with 500 nM DEX. P, Q Tet3 and LOX mRNA expression in osteoclasts in the 500 nM DEX group with or without RU486. Mean ± S.E.M., n = 3 per group for Bisulfite Sequencing PCR, western blotting, ChIP-PCR, and Co-IP, n = 8 per group for mRNA expression in vivo, n = 6 per group for mRNA expression in vitro. ^*P < 0.05, ^**P < 0.01 vs. Control in vivo or 0 nM DEX in vitro. ^##P < 0.01 vs. 500 nM DEX. PDE: prenatal dexamethasone exposure; LOX: lysyl oxidase; GR: glucocorticoid receptor; ERβ: estrogen receptor β; DEX: dexamethasone; Dnmt: DNA methyltransferase; Tet: ten-eleven translocation protein; SDHA: succinate dehydrogenase; TBP: TATA box binding protein; GAPDH: glyceraldehyde-3-phosphate dehydrogenase