Anti β actin antibody ac 15 a5441

Culture media (DMEM), fetal bovine serum (FBS) and penicillin/streptomycin (P/S) were obtained from Gibco (Grand Island, NY). Anti-p300 (N-15; sc-584) and anti-SP1 (PEP2; sc-59) antibodies were obtained from Santa Cruz Biochemistry (Santa Cruz, CA). Anti-Caspase3 (8G10; #9662), anti-cleaved Caspase3 (5A1E; #9664), anti-Caspase8 (8G10; #9662), anti-Caspase9 (8G10; #9662), anti-cleaved PARP (D64E10; #5625), anti-γ-H2AX (20E3; #9718), anti-Acetyl Histone H3 (Lys9) (C5B11; #9649), anti-Acetyl Histone H3 (Lys27) (D5E4; #8173), and anti-Histone H3 (D1H2; #4499) antibodies were obtained from Cell Signaling Technology (Danvers, MA). Anti-β-actin antibody (AC-15; A5441) was obtained from Sigma-Aldrich (St. Louis, MO). Gemcitabine hydrochloride (G6423) and the p300 inhibitor C646 (SML0002) were obtained from Sigma-Aldrich.

Antibodies were obtained from the following commercial sources: anti-β-actin antibody (AC-15, A5441) was purchased from Sigma-Aldrich; the anti-caspase-3 (#9662), anti-caspase-9 (#9502), anti-caspase-8 (#9746), anti-PARP (#9542), anti-c-Myc (#5605), anti-HSP90 (#4874), anti-Smad4 (#9515) and anti-H3 (#9715) antibodies were purchased from Cell Signaling Technology (Tokyo, Japan); the anti-phospho-Smad1/Smad5/Smad8 antibodies recognizing phosphorylated Ser463/465 were purchased from Millipore (MA, USA); and the anti-phospho-Ser62-c-Myc (ab51156) and anti-phospho-Thr58-c-Myc (ab135558) antibodies were purchased from Abcam (Cambridge, MA). After treatment, the cells were washed once with cold PBS, the cell pellets were treated with RIPA buffer, and the total protein was isolated. Protein (10 μg) was resolved in polyacrylamide gels and transferred onto polyvinylidene difluoride (PVDF) membranes (Millipore). The membranes were probed with the primary antibody described above followed by a horseradish peroxidase-linked secondary antibody (GE Healthcare Japan). The signals were detected using Chemi-Lumi One (Nacalai Tesque) with Image Quant LAS 4000 mini software (GE Healthcare, Buckinghamshire, UK). The band density was analyzed using the NIH ImageJ software.