Fig 2: Confirmation of protein sensitivity to changed glutathione redox ratios. A, MS-CETSA signals of PRDX1 and HAT1 proteins in LG-ITRR experiment. B, Western blotting-based CETSA validation of thermal stabilization of PRDX1 and HAT1 proteins in response to oxidation. C, SDS-PAGE analysis of PRDX1 and HAT1 proteins. HepG2 whole cell lysates (left and right panel) and HAT1 recombinant proteins (middle panel) were separated on SDS-PAGE under reducing (with DTT) or non-reducing (without DTT) conditions followed by western blotting detection. Arrows indicate protein monomers. The upper bands indicate dimer and oligomer formation of PRDX1 and HAT1 proteins upon GSSG addition, and inhibition in the presence of GSH.

Fig 3: Identification of the chromatin localization patterns of key histone acetyl transferases (HATs) and HDACs during mitosis.(A, B, C) Immunofluorescence of HeLa-S3 cells enriched for mitotic stages, stained for DNA (blue) and for various HDACs (A), sirtuins (B), and HATs (C) (green). Representative pictures from the major mitosis stages and interphase are shown, along with a quantification of at least 20 cells from each stage. Intensity values represent the mean ± SEM. Note that only HDAC3, P300, and HAT-1 are retained on the mitotic chromatin along all mitotic stages, whereas the other proteins show partial retention at prophase. (D) All factors (beside HDAC3, P300, and HAT1) show significant reduction at metaphase (P < 10-16, FDR-corrected t test) (D) Immunoblots showing the abundance of the indicated chromatin modifiers in the chromatin fraction of interphase (L1) and mitotic (L2) HeLa-S3 cells. Total histone H3 serves as loading control. Scale bars: 5 µm.Source data are available for this figure.

Fig 4: Involvement of HDAC3 in kcnq1ot1-induced downregulation of ABCA1 expression.A, B THP-1 macrophages were transfected with PBS, LV-NC or LV-kcnq1ot1 for 72 h. The expressions of LXRa, HDAC1, HDAC3, HDAC5, and HAT1 were determined by qRT-PCR and western blot (n = 3). C, D Evaluation of HDAC3 expression in the aorta from apoE-/- mice (n = 10). E, F Detection of HDAC3 expression in MPMs from apoE-/- mice. G THP-1 macrophages were transfected with scrambled siRNA or HDAC3 siRNA for 24 h, and the cell lysates were immunoblotted with indicated antibodies (n = 3). H–J THP-1 macrophages were transfected with HDAC3 siRNA for 24 h, followed by treatment with LV-kcnq1ot1 for another 72 h (n = 3). H, I The mRNA and protein levels of ABCA1 were detected by qRT-PCR and western blot, respectively. J Detection of cholesterol efflux to apoA-I using a liquid scintillation counter. Data are represented as mean ± SD. **P < 0.01, ***P < 0.001 vs. control group; ##P < 0.01, ###P < 0.001 vs. LV-kcnq1ot1 group; ns not significant.