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. 2020 May 20;10:8314. doi: 10.1038/s41598-020-65415-0

Figure 4.

Figure 4

Metformin increased histone acetylation and decreased DNMT1 and PGC-1α promoter methylation in human placental explants, concomitant with mitochondrial biogenesis signaling. Male placental explants from diabetic patients (n = 4 patients) were treated with metformin (1 mg/ml) or vehicle (H2O) for 18 hours in culture (plated in 4 wells in each group). (A,B) Levels of H3K27ac (A) and DNMT1 (B) were determined by Western blot analysis. Representation blots and bar graph of average fold changes in all male placental explants (n = 4 explants from different patients) were shown. (C,D) Placental explants with metformin treatment or vehicle controls were subjected for DNA methylation analysis and RNA extraction, and average of PGC-1α methylation of promoter b (C) and PGC-1α mRNA expression (D) in each individual placental explant were determined by RT-PCR (n = 3 explants from different patients). (E) Representation blots of P-ACC and P-AMPKα were shown. Mean ± SD, *P < 0.05 by paired T-test; #P < 0.05 by one-tail paired T-test. Full-length blots are provided in the Supplementary information.