Lymphocyte separation medium ficoll

HEK293, HCC38 and CaSki cell lines were purchased from American Type Culture Collection (ATCC). Cells were maintained in RPMI-1640 or DMEM culture media (Gibco, Life Technologies, USA) supplemented with 10% (v/v) fetal bovine serum (FBS) (HyClone Laboratories, USA). Normal human AB serum was acquired from Gibco, Life Technologies (USA). Anti-CD3 antibody, rhIL-2, rhTNF-γ, rhGM-CSF, rhIL-4 and TNF-α were purchased from Peprotech Inc. Lymphocyte separation medium Ficoll was purchased from GE Healthcare (USA). Mouse anti-human monoclonal antibodies of CD3 (FITC labeled) and CD8 (PE labeled) were purchased from eBioscience Co, Ltd. Anti-human PD1 and PDL1 monoclonal antibodies were purchased from BioLegend (San Diego, CA). Mouse anti-human monoclonal antibodies of CD80-PE, CD83-PE, CD86-PE, CD1a-FITC, and CD40-FITC were all purchased from Santa Cruz Co, Ltd. Female BALB/c nude mice (8-week-old, weight 16–22g) raised under SPF circumstance were acquired from the Guangzhou Traditional Chinese Medicine University. All animal studies were conducted in accordance with the Guide for the Care and Use of Laboratory Animal in accordance with the research protocols approved by Wuhan No.1 Hospital Animal Facility and Use Committee. PBMCs were generated from healthy donors with informed consent.

Chidamide was obtained from Shenzhen Chipscreen and prepared into 2 mM stock solution using Dimethyl sulfoxide (DMSO). The solution was preserved at −20 °C and thawed prior to use. RIPM1640 medium was purchased from Hyclone (USA). Fetal bovine serum (FBS) was purchased from Gibco (USA). CCK-8 was purchased from Dojindo (Japan). Annexin V-FITC apoptosis detection kit was manufactured by Dojindo. Anti-histone H3 antibody, anti-acetyl-histone H3 antibody, anti-p44/42MAPK (ERK1/2) antibody, and anti-Pp44/42MAPK (ERK1/2) antibody were purchased from Cell Signaling (USA). Anti-AML1/RHD antibody was purchased from Calbiochem (USA). Anti-C-KIT antibody, β-acting, and HRP-labeled secondary antibodies were purchased from Santa Cruz (USA). Lymphocyte separation medium (Ficoll) was purchased from GE Healthcare (USA). Trizol was purchased from Ambion (USA). Reverse transcription kit was purchased from Promega (USA). KAPA SYBR FAST q-PCR Master Kit was purchased from KAPA (USA).