Fig 2: P300/sirt7 is the key regulator of ginsenoside rb2 on sf3a2 (K10) acetylation. (A-B) The effect of NAM, TSA, A-485 or anacardic acid combined with ginsenoside Rb2 on the binding of SF3A2 with acetylated-lysine was observed by co-IP analysis. (C-D) After the knockdown with RNA interferences targeting different KATs and KDACs, the SF3A2 acetylation was observed in OGD/R-stress H9c2 cells. (E) After Rb2 incubation or p300, SIRT7 shRNA knockdown prior to OGD/R injury, the MMP were analyzed by FCM. (F-G) After shRNA knockdown for p300 or SIRT7, the effect of Rb2 on the level of SF3A2 (K10) acetylation was analyzed and quantified by Western blot in H9c2 cells. *P < 0.05, **P < 0.01 and ***P < 0.001. TSA: trichostatin A, NAM: nicotinamide.

Fig 3: H3K27ac mediated chromatin reorganization during aging.(a) Measurement of H3K27ac levels in Rag2−/− young (blue) and old (red) pro-B cells via flow cytometry. The IgG isotype controls are shown in dotted lines. The dot plot on the bottom indicates the median fluorescence intensities (MFI) of H3K27ac levels in young and old pro-B cells. Data are presented as mean ± SEM. Unpaired two-sided t test was used, with the P-value indicated. (b) Age-selective p300 ChIP-Seq binding sites were identified in young and old pro-B cells. The analysis and cutoffs used were consistent with those in Fig. 4a. (c) Overlaps of young selective H3K27ac and p300 peaks.(d) Age-selective Brg1 ChIP-Seq binding sites were identified in young and old pro-B cells. The analysis and cutoffs used were consistent with those in Fig. 4a. (e) H3K27ac HiChIP heatmaps of the Ebf1 locus in young (upper) and old (lower) pro-B cells. The visualization elements are similar to that shown in Fig. 4h. Source data

Fig 4: The analysis for the binding ability of different ginsenoside monomers on p300 protein. (A) Biophysical analysis of the interaction between p300/SIRT7 and ginsenoside Rb2 using a cellular thermal shift assay. (B) SPR analysis of ginsenoside Rb2 and A-485 binding to p300 protein and KD value was calculated. (C) The inhibitory effect of ginsenosides Rb2, Ro, Rg3, 20S-(PPT), or A-485 on p300 activity were assessed with a commercial kit.

Fig 5: MLF1 recruits EP300 to target promoters. (A) Venn diagram of MLF1, EP300 and H3K27ac-occupied genes. (B, C) CUT & Tag reveals the distribution changes of MLF1, EP300 and H3K27ac in the genome (B) and gene subsets List1 (C), following MLF1 knockdown. (D–F) Illustration of CUT & Tag peaks at the promoter regions of genes related to senescence (IL1B; D and p21; E) and hypertrophic markers (NPPA and NPPB; F) after MLF1 knockdown in AC16 cells. (G) Scatter plot of transcriptomic distribution in AC16 cells after MLF1 or EP300 knockdown. (H) Venn diagram of upregulated or downregulated DEGs in both MLF1 knockdown and EP300 gene silencing. (I) Heatmaps showing the DEGs that change concordantly following MLF1 and EP300 knockdown. (J) Functional enrichment of the 390 DEGs downregulated, following the knockdown of both MLF1 and EP300. (K) Overlapping relationship of upregulated genes after H2O2 treatment and downregulated genes after both MLF1 and EP300 knockdown. (L) Heatmaps display the 35 hub genes that are induced by H2O2 treatment and subsequently downregulated following the knockdown of both MLF1 and EP300. (M) Representative SA-β-Gal staining images (left) and quantification (right) showing the impact of the IL1B knockdown on the enhancement of SA-β-Gal staining after Adenovirus-mediated MLF1 overexpression in H2O2-treated AC16 cells. Scale bar, 100 μm. n = 5. *P< 0.05, ***P< 0.001 versus siNeg. ##P< 0.01, ###P< 0.001 versus Control.