Mouse anti β actin antibody

Protein samples were prepared by tissue lysate and BCA protein assay. Fifteen microliter protein of each sample was loaded and separated by 10% sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) and transferred onto polyvinylidene fluoride (PVDF) membranes (IPVH00010, Merck Millipore, Germany). The membranes were washed with 5% nonfat milk to block the non‐specific binding sites and then treated with primary antibodies: rabbit anti‐pad4 antibody, rabbit anti‐histone H3 (citrulline R2) antibody, mouse anti‐β‐actin antibody (M20011, Abmart, China) overnight. The secondary antibody was incubated with the membranes for 2 h. Signals were detected and exposed using LumiGLO^® Reagent (#7003, Cell Signaling Technology, USA).

Four human leukemia cell lines, K562, HL-60, U937 and THP1, were obtained from the American Type Culture Collection (Manassas, VA, USA). ATO was purchased from Beijing SL Pharmaceutical Co., Ltd. (Beijing, China) and the Cell Counting kit-8 (CCK-8) was purchased from Dojindo Laboratories (Kumamoto, Japan). An Annexin V-fluorescein isothiocyanate (AV) Apoptosis Detection kit and TRIzol reagent were purchased from Invitrogen Life Technologies (Carlsbad, CA, USA). A first-strand cDNA Quantscript RT kit and Taq DNA Polymerase were purchased from Tiangen Biotech Co., Ltd. (Beijing, China). Rabbit anti-EVI-1, anti-JNK, anti-phosphorylated-JNK (p-JNK), anti-Bax, anti-Bcl-2 and anti-B-cell lymphoma-extra large (Bcl-xL) and mouse anti-caspase-3 antibodies were obtained from Cell Signaling Technology, Inc. (Danvers, MA, USA). Mouse anti-β-actin antibody was purchased from Abmart (Arlington, MA, USA). Horseradish peroxidase-conjugated secondary antibodies were purchased from Jackson ImmunoResearch Laboratories, Inc. (West Grove, PA, USA).