Anti p300

The immunoblot analysis was performed as described previously 35 (link). Briefly, tissues and cells were lysed in 1×RIPA buffer (Sigma-Aldrich, #R0278). Equal amounts of protein in each sample were loaded into a 10% SDS-PAGE gel. After transferring to a membrane and blocking with 5% milk, the proteins were probed with the following primary antibodies: anti-CtBP1 (BD Biosciences, San Jose, CA, USA, #612042), anti-CtBP2 (BD Biosciences, #612044), anti-CD31 (ThermoFisher Scientific, #PA5-16301), anti-CD55 (ThermoFisher Scientific, #PA5-82005), anti-CD68 (ThermoFisher Scientific, #MA5-13324), anti-GAPDH (Santa Cruz Biotechnology, Dallas, Texas, USA, #sc-365062), anti-Caspase-1 (Santa Cruz Biotechnology, #sc-56036), anti-Flag (Sigma-Aldrich, #SAB4200071), anti-Myc (Abcam, Cambridge, MA, USA, #ab9106), anti-p300 (Santa Cruz Biotechnology, #sc-585), anti-c-Jun (Sigma-Aldrich, #SAB4501606), anti-c-FOS (Sigma-Aldrich, #F7799), anti-p50 (ThermoFisher Scientific, #PA1-30409), anti-p65 (ThermoFisher Scientific, #14-6731-81), anti-IRF2 (Abcam, #ab3388), anti-STAT4 (Abcam, #ab68156), anti-NLRP3 (Abcam, #ab210491), anti-Il-1β (Abcam, #ab2105), anti-DNMT1 (Abcam, #ab13537), and anti-DNMT3A (Abcam, #ab2850). After probing with secondary antibodies, protein band signals were detected using a Pierce^TM ECL western blotting substrate (ThermoFisher Scientific, #32106).

Immunoprecipitation and imunoblotting were performed as described previously [26 (link)]. The reagents were as following: anti-β-catenin (BD Bioscience, Cat# 610153), anti-γ-catenin (BD Bioscience, Cat# 610253), anti-CBP (Santa Cruz, clone SC-369), anti-p300 (Santa Cruz, clone SC-584), anti-activated- β-catenin (Millipore, clone 8E7), anti-lamin A/C (Santa Cruz, SC-7293), NE-PER Nuclear extraction reagent (Pierce, Cat#78833), Protease inhibitor cocktail (Calbiochem, Cat#539137), Protein A-agarose (Roche, Cat#11134515001), Illustra microspin columns (GE Healthcare, Cat#27-3565-01), ECL Plus (GE Healthcare, Cat#RPN 2132), and Blue ultra autorad film (BioExpress, Cat# F9029-8X10).

As previously described^{17 (link)}, chromatin immunoprecipitation (ChIP) was performed using EZ-Magna ChIP A and EZ-Magna ChIP G Kits (Millipore, USA), following the manufacturer’s instructions. Briefly, ChIP grade antibodies were as follows: anti-histone H3 lysine 27 acetylation (H3K27ac; Thermo Fisher, USA), anti-histone H3 lysine 4 trimethylation (H3K4me3; Millipore, USA), anti-histone H3 lysine 27 trimethylation (H3K27me3; Millipore, USA), anti-P300 (Santa Cruz Biotechnology, USA), anti-CBP (Thermo Fisher, USA), anti-SMYD3 (Millipore, USA), and normal IgG (Millipore, USA). Immunoprecipitated DNA was amplified by qPCR, and normalized to the input DNA. Primer sequences for the promoter region of NUF2 are listed in Supplementary Table S1.

Chromatin immunoprecipitation (ChIP) assays were performed essentially as described before (Li et al., 2019b (link)). In brief, chromatin in control and treated cells were cross-linked with 1% formaldehyde. Cells were incubated in lysis buffer (150 mM NaCl, 25 mM Tris pH 7.5, 1% Triton X-100, 0.1% SDS, 0.5% deoxycholate) supplemented with protease inhibitor tablet and PMSF. DNA was fragmented into ∼200 bp pieces using a Branson 250 sonicator. Aliquots of lysates containing 200 μg of protein were used for each immunoprecipitation reaction with anti-BRG1 (Santa Cruz, sc-10768), anti-anti-acetyl H3 (Millipore, 06-599), anti-dimethyl H3K9 (Millipore, 17-648), anti-HIF-1α (Santa Cruz, sc-10790), anti-p300 (Santa Cruz, sc-585), anti-KDM3A (Abcam, ab-91252), or pre-immune IgG. For re-ChIP, immune complexes were eluted with the elution buffer (1% SDS, 100 mM NaCO₃), diluted with the re-ChIP buffer (1% Triton X-100, 2 mM EDTA, 150 mM NaCl, 20 mM Tris pH 8.1), and subject to immunoprecipitation with a second antibody of interest.