Journal: Journal of Translational Medicine

Article Title: Microbiota-associated metabolite pantothenic acid enhances skeletal muscle contusion repair via epigenetic regulation of macrophage M2 polarization

doi: 10.1186/s12967-026-08087-0

Figure Lengend Snippet: Pantothenic acid synergizes with IL-4 to drive M2 polarization by upregulating Arg1 expression through mediating H3K27ac. ( A ) Intracellular acetyl-CoA levels in each group of cells ( n = 6). ( B - C ) Relative protein expression levels of H3K27ac in each group of cells ( n = 6). ( D ) Enrichment of H3K27ac at the Arg1 gene promoter region in each group of cells ( n = 3). ( E ) Relative mRNA expression of Arg1 in each group of cells ( n = 7). ( F ) Enrichment of H3K27ac at the Arg1 gene promoter region in each group of mice ( n = 3). ( G ) Relative mRNA expression of Arg1 in each group of mice ( n = 6). ( H ) Intracellular acetyl-CoA levels in each group of cells ( n = 6). ( I - J ) Relative protein expression levels of H3K27ac in each group of cells ( n = 6). ( K - M ) Relative protein expression levels of CD206 ( L ) and Arg1 ( M ) in each group of cells ( n = 6). All data are presented as mean ± SEM. Statistical significance among multiple groups was determined by one-way ANOVA. * P < 0.05, ** P < 0.01, and *** P < 0.001

Article Snippet: Chromatin was immunoprecipitated overnight at 4°C with an anti-H3K27ac antibody (Huabio, HA600047 ) or normal Rabbit IgG.

Techniques: Expressing

Journal: Molecular Metabolism

Article Title: Nutrient-driven histone acetylation underlies energy storage and mobilization

doi: 10.1016/j.molmet.2026.102344

Figure Lengend Snippet: BHB-mediated HDAC inhibition suppresses lipolysis in the WAT. (A) The body weight loss and the blood BHB levels of the mice underwent starvation at the indicated time points ( n = 5). (B) Pearson correlation analysis of the blood BHB levels and body weight loss rate. (C and E) Western blotting assay of H3K27ac in the white adipose tissue (WAT) of mice underwent fasting (C) or re-feeding (E) at the indicated time points (left); the quantification of the bands (right) ( n = 5). (D and F) The blood glucose and BHB levels of mice underwent fasting (D) or re-feeding (F) at the indicated time points ( n = 5). (G and H) The medium FFA levels of the adipocytes differentiated from 3T3-L1 (G) or C3H10T1/2 (H) with indicated treatment ( n = 3–4). JQ35 (1 μM), BHB (20 mM), and Merck60 (1 μM) pretreated for 4 h, followed by isoproterenol (ISO, 10 μM) treatment for 3 h. (I) The medium FFA levels of WAT pretreated with BHB ± JQ35, followed by 2 h of ISO treatment. ( n = 3). (J) qPCR assays of lipolytic genes ( Hsl , Atgl ) of the adipocytes differentiated from C3H10T1/2 with the indicated treatment ( n = 3). (K and L) The body weight (K) and the blood BHB levels (L) of mice underwent fasting or fasting plus Merck60 ( n = 5). (M and N) Pearson correlation analysis of the blood BHB levels with core body temperature (M) or heart rate (N). Data are shown as the mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001 by unpaired two-tailed Student’s t-test (L), one-way (C, E, and G-J), or two-way ANOVA (D, F, and K) followed with Bonferroni’s multiple comparisons test.

Article Snippet: ChIP was performed using a SimpleChIP® Plus Enzymatic Chromatin IP Kit (9005; Cell Signaling Technology) with antibodies against H3K27ac (5 μg/ChIP, activemotif, 91193), BRD4 (5 μg per ChIP, Cell Signaling Technology, 83375S), or normal rabbit IgG as control according to the manufacturer’s procedures.

Techniques: Inhibition, Western Blot, Two Tailed Test

Journal: Molecular Metabolism

Article Title: Nutrient-driven histone acetylation underlies energy storage and mobilization

doi: 10.1016/j.molmet.2026.102344

Figure Lengend Snippet: BHB-mediated HDAC inhibition shifts hepatic anabolism toward lipid oxidation. (A) Experimental design. (B) Serum gross appearance and the FFA levels ( n = 5). (C) Body weight loss of mice with the indicated treatment ( n = 5). (D and E) ChIP-seq data displaying H3K27ac (D) or BRD4 (E) occupancy at two subsets of genes: H3K27ac and BRD4 enriched at the genomic loci of fasting-induced transcripts (energy mobilization associated genes, such as Cyp4a14), but dislodged from those loci of fasting-suppressed transcripts (energy storage-associated genes, such as Pcsk9). (ref. to Hsieh et al., 2022, Mol Cell for detail method of analysis). (F and G) GO enrichment analysis of the fasting-induced H3K27ac (F) and BRD4 (G) peak-associated genes. The reads number of the ChIP-seq data for H3K27ac (log2FC > 1 and P < 0.01) or BRD4 (log2FC > 1.2 and P < 0.01) were included as the fasting-induced peaks. (H and I) Gene tracks display that fasting induced the occupancy of H3K27ac and BRD4 at the genomic loci of energy mobilization-associated genes (Cyp4a14, Pck1) (H), but reduced at those loci of energy storage-associated genes (Pcsk9, Srebf1) (I). (J) The ratio of the liver to body weight (upper panel) and the liver triglycerides levels (lower panel) of mice with the indicated treatment ( n = 5). (K-O) Body weight loss data (K), serum gross appearance and FFA levels (L), the blood glucose and BHB levels (M), the ratio of liver to body weight (N) and Liver triglycerides (O)of Brd 4 -flox or Brd4 hKO mice subjected to 24 h of fasting ( n = 5). Data are shown as the mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001 by unpaired two-tailed Student’s t-test (K-O) or one-way ANOVA followed with Bonferroni’s multiple comparisons test (B, C, and J).

Article Snippet: ChIP was performed using a SimpleChIP® Plus Enzymatic Chromatin IP Kit (9005; Cell Signaling Technology) with antibodies against H3K27ac (5 μg/ChIP, activemotif, 91193), BRD4 (5 μg per ChIP, Cell Signaling Technology, 83375S), or normal rabbit IgG as control according to the manufacturer’s procedures.

Techniques: Inhibition, ChIP-sequencing, Two Tailed Test

Journal: Molecular Metabolism

Article Title: Nutrient-driven histone acetylation underlies energy storage and mobilization

doi: 10.1016/j.molmet.2026.102344

Figure Lengend Snippet: BHB-mediated HDAC inhibition sustains BAT fasting biology. (A) Western blotting assay of H3K27ac in the BAT of mice with or without 24 h of fasting (upper panel); the quantification of the bands (lower panel) ( n = 4). (B and C) GO (B) and KEGG (C) enrichment analysis of fasting-induced peak-associated genes. The reads number of the CUT&Tag-seq data for H3K27ac (log2FC > 0.8 and P < 0.05) were included as the fasting-induced peaks. (D) Gene tracks display that fasting induced the occupancy of H3K27ac at the genomic loci of FAO associated genes, but not that of Ucp1 . (E) Experimental design. (F-H) The core body temperature of the mice subjected to the indicated treatment ( n = 4–5). (I) Glucose administration enhanced fasting-caused hypoglycemia (left) and rescued JQ35-suppressed thermogenesis (right) ( n = 4–5). (J and K) The core body temperature, blood BHB and glucose levels of Brd 4 -flox or Brd 4 -hKO mice subjected to 24 h of fasting (J) ( n = 5) or fed with ketogenic diet (K) ( n = 4–5). Data are shown as the mean ± SEM. ∗ P < 0.05, ∗∗∗∗ P < 0.0001 by unpaired two-tailed Student’s t-test (A, I, J, and K) or two-way ANOVA followed with Bonferroni’s multiple comparisons test (F-I).

Article Snippet: ChIP was performed using a SimpleChIP® Plus Enzymatic Chromatin IP Kit (9005; Cell Signaling Technology) with antibodies against H3K27ac (5 μg/ChIP, activemotif, 91193), BRD4 (5 μg per ChIP, Cell Signaling Technology, 83375S), or normal rabbit IgG as control according to the manufacturer’s procedures.

Techniques: Inhibition, Western Blot, Two Tailed Test

Journal: Molecular Metabolism

Article Title: Nutrient-driven histone acetylation underlies energy storage and mobilization

doi: 10.1016/j.molmet.2026.102344

Figure Lengend Snippet: BHB-mediated HDAC inhibition promotes renal gluconeogenesis and anti-oxidation ability. (A) Western blotting assay of H3K27ac in the kidney of mice with or without 24 h of fasting (upper panel); the quantification of the bands (lower panel) ( n = 5–7). (B and C) Gene tracks displayed the differential occupancy of H3K27ac at the genomic loci of Pck1 (massively increase) and G6pc (slightly increase) (B), which matches with the differential transcriptional output of the genes (C) ( n = 5). (D and E) Western blotting (D) and qPCR (E) assay of HK2 and Caco-2 cells treated with LY294002 (1 μM), BHB (20 mM), or Merck60 (1 μM) for 24 h ( n = 4). (F) Transcription factor motif enrichment analysis of the kidney ChIP-seq datasets. (G and H) Gene tracks displayed the occupancy of H3K27ac at the genomic loci of anti-oxidative genes ( Cat , Gstt2 , Gpx3 , Hnf4a ) (G), and JQ35 treatment suppressed the expression of these genes (H) ( n = 5). Data are shown as the mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001 by unpaired two-tailed Student’s t-test (A, C, and H) or one-way ANOVA followed with Bonferroni’s multiple comparisons test (E).

Article Snippet: ChIP was performed using a SimpleChIP® Plus Enzymatic Chromatin IP Kit (9005; Cell Signaling Technology) with antibodies against H3K27ac (5 μg/ChIP, activemotif, 91193), BRD4 (5 μg per ChIP, Cell Signaling Technology, 83375S), or normal rabbit IgG as control according to the manufacturer’s procedures.

Techniques: Inhibition, Western Blot, ChIP-sequencing, Expressing, Two Tailed Test

Journal: Molecular Metabolism

Article Title: Nutrient-driven histone acetylation underlies energy storage and mobilization

doi: 10.1016/j.molmet.2026.102344

Figure Lengend Snippet: Glucose-mediated histone acetylation promotes lipogenesis. (A-D) ChIP-seq data showed that re-feeding induced a significant redistribution of H3K27ac and BRD4 in the whole genome (A and B) with an increased occupancy at the genomic loci of lipids synthesis genes ( Srebf1 , Pcsk9 ) (C) but a decreased occupancy at those of fatty acids catabolic ( Cyp4a14 ) and gluconeogenic genes ( Pck1 ) (D). (E and F) qPCR assay showed that re-feeding induced the expression of glucose utilization ( Gck ) and lipid synthesis genes ( Pcsk9 , Hmgcr , Dgat1 , Srebf1 ) in the liver (E), while the expression was suppressed by JQ35 treatment (F) ( n = 4). (G and H) Brd 4 -flox or Brd 4 -hKO mice housed in a thermoneutral environment (30°C) feeding with high-fat diet. Hepatic Brd4 knockout suppressed body weight gain (G) without influence food intake (H) ( n = 5–6). (I-L) ITT assays (I) , liver weight (J), the liver gross appearance, HE and Oil Red O staining (K), and TG levels (L) of Brd 4 -flox or Brd 4 -hKO mice that were subjected to HFD feeding with housing at 30°C ( n = 5–6). (M) qPCR assay of lipids anabolism- ( Cd36 , Pparg , Dgat1 ) and VLDLs secretion/metabolism- ( Mttp , Apoc3 ) associated genes in the liver of Brd 4 -flox or Brd 4 -hKO mice ( n = 5–6). (N) The serum ALT levels were lower in Brd 4 -hKO than that of control mice ( n = 5–6). (O-Q) WAT weight (O) and serum TG (P) and FFA (Q) levels of Brd 4 -flox or Brd 4 -hKO mice under HFD feeding ( n = 5–6). Data are shown as the mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001 by unpaired two-tailed Student’s t-test (E, F, H, J, and L-Q) or two-way ANOVA followed with Bonferroni’s multiple comparisons test (G and I).

Article Snippet: ChIP was performed using a SimpleChIP® Plus Enzymatic Chromatin IP Kit (9005; Cell Signaling Technology) with antibodies against H3K27ac (5 μg/ChIP, activemotif, 91193), BRD4 (5 μg per ChIP, Cell Signaling Technology, 83375S), or normal rabbit IgG as control according to the manufacturer’s procedures.

Techniques: ChIP-sequencing, Expressing, Knock-Out, Staining, Control, Two Tailed Test

Journal: Molecular Metabolism

Article Title: Nutrient-driven histone acetylation underlies energy storage and mobilization

doi: 10.1016/j.molmet.2026.102344

Figure Lengend Snippet: ACSS2 is required for in vitro and in vivo lipogenesis. (A and B) 3T3-L1 preadipocytes were pretreated with ACLY (5 μM) or ACSS2 (5 μM) inhibitors, followed by adipogenic induction. Oil Red O staining (day 8 after adipogenic induction) (A, n = 3) and western blotting assay (day 1 after adipogenic induction) (B, n = 3) were performed at indicated times. (C and D) 3T3-L1 preadipocytes (at the cell confluence of 50–60%) were transfected with Acly or Acss2 siRNA. Two days after the cells reaching at 100% confluence, they were subjected to adipogenic induction. Oil Red O staining (day 8 after adipogenic induction) (C, n = 3) and western blotting assays (day 1 after adipogenic induction) (D, n = 3) were performed at indicated times. (E and F) Stromal vascular fractions (SVFs) were isolated from Acss2 fl/fl subcutaneous adipose depots and the cells were infected with Ad-GFP or Ad-Cre, followed by adipogenic induction. Oil Red O staining (day 8 after adipogenic induction) (E, n = 3) and western blotting assays (day 2 after adipogenic induction) (F, n = 3) were performed at indicated times. (G and H) qPCR assay of glucose metabolism ( G6pd , Me1 , Pklr ) (G) and fatty acid synthesis genes ( Fasn , Chrebpb , Acaca ) (H) in the liver of mice fed with fructose and glucose water and treated with ACLYi (50 mg/kg, i.p., bid), ACSS2i (40 mg/kg, i.p., bid), or both inhibitors ( n = 4–5). (I) Western blotting assay of ACSS2 in the liver of Acss 2 -flox or Acss 2 -hKO mice (upper panel); the quantification of the bands (lower panel) ( n = 4). (J) GO enrichment analysis of down-regulated genes in the liver of Acss 2 -hKO versus Acss 2 -flox mice subjected to 24 h fructose and glucose water exposure. (K) qPCR assay of Acss2 and fatty acid synthesis genes ( Acaca , Chrebpb , Fasn ) in the liver of Acss2 flox and Acss 2 -hKO mice subjected to 24 h of fructose and glucose water exposure ( n = 4). (L) ChIP was performed using anti-H3K27ac antibody or control IgG, and DNA was amplified with specific primers to detect the target genes ( Pklr and Mlxipl ) ( n = 4). (M) The ratio of liver to body weight (left) and the liver TG levels (right) of Acss 2 -flox or Acss 2 -hKO mice subjected to 24 h of fructose and glucose water exposure ( n = 4). Data are shown as the mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001 by unpaired two-tailed Student’s t-test (I, K, and M) or one-way ANOVA followed with Bonferroni’s multiple comparisons test (G, H, and L).

Article Snippet: ChIP was performed using a SimpleChIP® Plus Enzymatic Chromatin IP Kit (9005; Cell Signaling Technology) with antibodies against H3K27ac (5 μg/ChIP, activemotif, 91193), BRD4 (5 μg per ChIP, Cell Signaling Technology, 83375S), or normal rabbit IgG as control according to the manufacturer’s procedures.

Techniques: In Vitro, In Vivo, Staining, Western Blot, Transfection, Isolation, Infection, Control, Amplification, Two Tailed Test

Journal: Journal of Advanced Research

Article Title: The GOLM1-ACLY pathway regulates macrophage-secreted EFEMP1 via H3K27ac modifications to drive tumor progression

doi: 10.1016/j.jare.2025.07.003

Figure Lengend Snippet: Nucleic P-ACLY within Golm1 -knockout macrophage regulates Efemp1 expression through H3K27 acetylation (A) Total histone H3 acetylation levels in Thp1 nc and Thp1 −/− cells determined by Histone H3 acetylation detection Kit. n = 9 replicated wells. (B-C) WB detecting (B) and quantitative analysis (C) of the protein levels of various acetylated histone H3. n = 3 independent experiments. (D) PCA plot of CUT&Tag-seq data of experiments with Thp1 nc and Thp1 −/− cells. n = 3 samples/group. (E) Pie chart showing the distribution of H3K27ac peaks across annotated genomic regions in Thp1 nc and Thp1 −/− cells. (F) Heatmap showing the genomic occupancy of H3K27ac from −3 kb flanking TSS to + 3 kb flanking TES in Thp1 nc or Thp1 −/− cells. The x-axis represents the position of reads relative to the TSS, and the y-axis represents the read density. (G) CUT&Tag-seq volcano plot of different gene peaks between Thp1 nc and Thp1 −/− cells. (H) RNA-seq volcano plot of DEGs between BMDMs wt and BMDMs mko cells. n = 3 mice/group. (I) Overlap of differentially expressed genes identified by RNA-seq and CUT&Tag-seq. Secreted proteins with statistically significant differences are listed below. (J) IGV snapshot of Thp1 nc and Thp1 −/− cells showing the H3K27ac CUT&Tag signals at the Efemp1 genomic loci. (K) H3K27ac occupancy in the genome of Efemp1 analysed by ChIP-qPCR. n = 4 duplicates/group. (L) Relative expression of Efemp1 genes in Thp1 nc and Thp1 −/− cells determined by qPCR. n = 6 duplicates/group. The data are presented as the means ± SEMs. The results presented in (A, C, K, and L) were analyzed via two-tailed Student's t test. ns, not statistically significant. **p < 0.01; ***p < 0.001.

Article Snippet: The Bead-bound cells were resuspended and sequentially incubated with anti-H3K27ac primary antibody (8173S, Cell Signaling Technology, USA) and secondary antibody (TransGen Biotech, China).

Techniques: Knock-Out, Expressing, RNA Sequencing, ChIP-qPCR, Two Tailed Test