S of DNMT1 or RBBP7 and subjected to PSS. n = 3 independent experiments. P sirtuininhibitor 0.05.Author Manuscript Author ManuscriptSci Signal. Author manuscript; obtainable in PMC 2018 February 28.Marin et al.PageAuthor Manuscript Author ManuscriptFig. three. AMPK activation decreased promoter methylation of PGC-1, NRF1, NRF2, Tfam, UCP2, and UCP3 genes(A to C) Methylation-specific quantitative polymerase chain reaction (qPCR) analysis of promoter methylation in AICAR- or metformin-treated AMPK+/+ and AMPK-/- MEFs in comparison to nontreated AMPK+/+ group (A); HUVECs transfected with DNMT1-WT, DNMT1-S730A, or DNMT1-S730D (B); or HUVECs transfected with RBBP7-WT, RBBP7-S314A, or RBBP7-S314D (C). Cells have been treated as indicated. n = four independent experiments for (A) to (C). (D) Promoter methylation status in HUVECs transfected with DNMT1-WT, DNMT1-S730A, or DNMT1-S730D and subjected to PSS. n = 3 independent experiments. P sirtuininhibitor 0.05. FC, fold transform.Author Manuscript Author ManuscriptSci Signal. Author manuscript; readily available in PMC 2018 February 28.Marin et al.PageAuthor Manuscript Author ManuscriptFig.RSPO1/R-spondin-1 Protein Molecular Weight four. AMPK improved HAT1 activityHAT1 activity was assessed in cells treated with AICAR or metformin or left untreated. (A) AMPK+/+ and AMPK-/- MEFs. (B) AMPK-/- MEFs had been infected with Ad-AMPK-CA or Ad-AMPK-DN AMPK (50 MOI) before remedy. n = three independent experiments for (A) and (B). (C) HAT1 activity in HUVECs transfected with control, AMPK, HAT1, or RBBP7 siRNA prior to the indicated remedy. n = four independent experiments. (D) HAT1 activity in cells transfected with handle, AMPK, HAT1, or RBBP7 siRNA then subjected to PSS. n = three independent experiments. (E and F) HUVECs had been transfected together with the indicated forms of HAT1 or RBBP7 and treated as indicated. n = 4 independent experiments for (E) and n = 3 independent experiments for (F).IL-18, Human (HEK293, His) P sirtuininhibitor 0.PMID:23618405 05.Author Manuscript Author ManuscriptSci Signal. Author manuscript; available in PMC 2018 February 28.Marin et al.PageAuthor Manuscript Author ManuscriptFig. five. AMPK activation decreased nucleosomal compaction of PGC-1, NRF1, NRF2, Tfam, UCP2, and UCP3 genes(A) H4K5 acetylation in AMPK+/+ or AMPK-/- MEFs treated with or devoid of AICAR or metformin. n = 4 independent experiments. (B) H4K5 acetylation in HUVECs expressing indicated forms of HAT1 and treated as indicated. n = 4 independent experiments. (C) H4K5 acetylation in HUVECs expressing the indicated kinds of RBBP7 and treated as indicated. n = 4 independent experiments. (D) H4K5 acetylation in cells expressing the indicated forms of HAT1 then subjected to PSS. n = 3 independent experiments. (E to G) FAIRE analysis in HUVECs transfected using the indicated types of DNMT1, RBBP7, or HAT1 and treated as indicated. n = four independent experiments. P sirtuininhibitor 0.05.Author Manuscript Author ManuscriptSci Signal. Author manuscript; accessible in PMC 2018 February 28.Marin et al.PageAuthor Manuscript Author ManuscriptFig. six. AMPK activation elevated mitochondrial biogenesis and functionHUVECs expressing the indicated forms of DNMT1, RBBP7, or HAT1 were treated with AICAR or left untreated. (A) JC-1 staining. n = six independent experiments. (B) MitoTracker staining. n = 6 independent experiments. (C) Quantification of MitoTracker staining of six fields. n = 6 independent experiments. (D) Mitochondrial DNA (mtDNA) abundance. (E to H) Activity of citrate synthase (E), complicated I (F), complex IV (G), and complex V (H). (I) ATP ab.
