Histone h3 acetyl k18

Pancreatic cancer cells were cultured with four concentrations of JYQ-42 (0, 10, 20, and 40 μmol/L) for 24 or 48 h. After treatment, the cells were collected and lysed with 1 × SDS loading buffer. Total proteins were resolved on 12% SDS-PAGE, transferred to PVDF membranes. 50 mmol/L Tris-HCl (pH 7.5) with 100 mmol/L NaCl, 0.5% (v/v) Tween-20 and 5% nonfat milk was used for blocking. 50 mmol/L Tris-HCl (pH 7.5) with 100 mmol/L NaCl and 1% BSA was used to prepare primary and secondary antibodies. An Immobilon Western Chemiluminescent HRP Substrate Kit (Merck Millipore) was used for detection of the protein bands and blots were scanned with G: BOX Chemi system (Syngene, Cambridge, UK).
Antibodies to the following proteins were used in the Western blots: HRP-linked anti-rabbit IgG (Cell Signaling, 7074P2; 1:20,000); HRP-linked anti-mouse IgG (Cell Signaling, 7076P2; 1:20,000); SIRT6 (Cell Signaling, 12486; 1:2000); Histone H3 (Abcam, ab10799; 1:2000); Histone H3 (acetyl K9) (Abcam, ab32129; 1:1000); Histone H3 (acetyl K18) (Abcam, ab1191; 1:1000); Histone H3 (acetyl K56) (Active Motif, 39281; 1:1000); β-actin (Proteintech Group, HRP-60008; 1:5000).