Histone 3

Cells and tissues were lysed in RIPA (P0013B, Beyotime, China) supplemented with 1 mM PMSF (ST506, Beyotime, China) and 0.01 mM phosphatase inhibitors (P1260, Solarbio, China). Samples with 20 ug protein in each group were separated by 10% SDS-PAGE and electrotransferred to PVDF membranes (Millipore, USA). The primary antibodies were used as following: β-actin (1:2000, 20536-1-AP, Protein tech, China), EZH2 (1:1000, #5246, Cell Signaling Technology, USA), SUZ12 (1:1000, #3737, Cell signaling Technology, USA), EED (1:1000, 16818-1-AP, Protein tech, China), Histone 3 (1:2000, 17168-1-AP Protein tech, China), H3K27me3 (1:1000, #9733, Cell Signaling Technology, USA), and H3K27ac (1:1000, #8173, Cell Signaling Technology, USA). After incubated with horseradish peroxidase-conjugated secondary antibody (1:2000, #7074, Cell Signaling Technology, USA), the blots were visualized with chemiluminescent substrate (US Everbright Inc., China) by Image Lab (Bio-Rad, USA).

Primary antibodies against NDUFS1 (no. 12444), NDUFA9 (no. 20312-1-AP), NDUFV1 (no. 11238), NDUFV2 (no. 5301), KU80 (no. 6389), KU70 (no. 10723), RAD51 (no. 14961), PARP-1 (no. 13371), PDP1 (no. 21176),PDK1 (no. 10026), β-actin (no. 60008), and Histone3 (no.17168) were obtained from Proteintech and γ-H2AX (no. AP0099), PDHA1 (no. A17432) were obtained from Abclonal and p-PDH (no.ab115343), HIF1a (no. ab179483) were obtained from Abcam, and Acetyl-Histone H3 (Lys9) (no. 9649 s), Acetyl-Histone H3 (Lys56) (no. 4243 s), were obtained from CST in this study.

After being treated with LPS, the total protein level of the BMDMs was collected using RIPA (Beyotime, China) and protease inhibitor (Roche, USA). Additionally, the cytoplasmic and nuclear protein fractions were extracted by using the ProteinExt Mammalian Nuclear and Cytoplasmic Protein Extraction Kit (TransGen Biotech, China). The levels of TRIM59 (Abcam, USA), p65 (CST, USA), p-p65 (CST, USA), IκB (CST, USA), p- IκB (CST, USA), IKKα (CST, USA), IKKβ (CST, USA), p-IKKα/β (CST, USA), ECSIT (ABclonal Biotechnology, China), MAP3K1 (ABclonal Biotechnology, China), AKT (CST, USA), p-AKT^Ser473 (CST, USA), p-AKT^Thr308 (CST, USA), PI3K (CST, USA), p-PI3K (CST, USA), GAPDH (Proteintech, China), and Histone-3 (H3, Proteintech, China) were measured using Western blot analysis.