For assessment of endogenous histone acylation, cells and embryos were lysed in the radioimmunoprecipitation assay (RIPA) buffer [150 mM NaCl, 1.0% NP-40, 0.5% sodium deoxycholate, 0.1% SDS, 50 mM tris-HCl (pH 8.0), and protease inhibitor mixture] for immunoblotting. Histone acylation was analyzed by immunoblotting with anti-H3 (Abcam, ab1791; 1:100,000), anti-H3K9ac (Abcam, ab10812; 1:1000), anti-H3K18ac (EMD Millipore, 07-354; 1:1000), anti-H3K23ac (EMD Millipore, 07-355; 1:10,000), anti-H3K23pr (PTM Biolabs, PTM-205; 1:1000), anti-H3K23cr (PTM Biolabs, PTM-519; 1:1000), anti-H3K23bu (PTM Biolabs, PTM-307; 1:1000), anti-H3K23me3 (Active Motif, 61499; 1:1000), anti-H3K14ac (EMD Millipore, 07-353; 1:1000), anti-H3K14pr (PTM Biolabs, PTM-211; 1:1000), anti-H3K14cr (PTM Biolabs, PTM-535; 1:1000), anti-H3K14hib (PTM Biolabs, PTM-881; 1:1000), anti-propionyllysine (PTM Biolabs, PTM-201; 1:1000), anti-crotonyllysine (PTM Biolabs, PTM-502; 1:1000), and anti-succinyllysine (PTM Biolabs, PTM-402; 1:1000) antibodies in 5% nonfat milk or bovine serum albumin in the TBST (Tris-buffered saline with Tween 20) buffer for overnight at 4°C. Goat anti-Rabbit IgG (H+L)-HRP (Thermo Fisher Scientific, 65-6120) and Goat anti-Mouse IgG (H+L)-HRP (Thermo Fisher Scientific, 62-6520) were used as the secondary antibodies at a dilution of 1:5000 for blotting for 1 hour at room temperature. After extensive washing four times with the TBST buffer (7 min each), blots were developed with the Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific, 32109).

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