P21 antibody

The EdU kit (Cat No: C10310-1) was purchased from RiboBio, Guangzhou, China, and the extreme ultrasensitive ECL chemiluminescence kit (Cat No: P0018FM) was purchased from Beyotime, China. Trypsin (Cat No: T1300), MTT reagent (Cat No: M8180), cell-grade dimethyl sulfoxide DMSO (Cat No: D8371) and Rainbow 180 Broad Spectrum Protein Marker (Cat No: PR1910) were purchased from Solarbio, Beijing, China. DMEM high glucose medium (Cat No: SH30243.01) was purchased from Gibco, New York, NY, USA, and 100 U penicillin–streptomycin solution (Cat No: SV30010) was purchased from Hyclone, Los Angeles, CA, USA. PVDF membranes, 5-fluorouracil (5-FU) and Entinostat (Ent) were obtained from MERCK, Darmstadt, Germany. Bcl2 antibody (Cat No: 68103-1-Ig), Caspase-3 antibody (Cat No: 66470-2-Ig), Cytochrome C (Cyt C) antibody (Cat No: 10993-1-AP), p53 antibody (Cat No: 60283-2-Ig), p21 antibody (Cat No: 10355-1-AP), CDK2 antibody (Cat No: 10122-1-AP), HDAC1 antibody (Cat No: 66085-1-Ig), HDAC2 antibody (Cat No: 67165-1-Ig), HDAC3 antibody (Cat No: 67151-1-Ig) and HDAC8 antibody (Cat No: 17548-1 AP) were purchased from Proteintech, Wuhan, China. H4 antibody (Cat No: PTM-1009), H3K18 antibody (Cat No: PTM-114), H3K27 antibody (Cat No: PTM-5010) and H3K9 antibody (Cat No: PTM-7094) were purchased from Jingjie PTM BioLab, Hangzhou, China.

Cells were cultured with vehicle or 2.5 μmol/L pentamidine in 6‐well plates (10⁶ cells/well) at 37°C for 24 hours and then lysed using RIPA Lysis and Extraction Buffer (Thermo Scientific, 89901) containing the protease inhibitor cocktail (Thermo Scientific, 87786). Proteins were then separated by SDS‐polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes. After blocked with 5% fat‐free milk for 1 hour at room temperature, the membranes were incubated with primary antibodies at 4°C overnight: P21 antibody (Proteintech, 10355‐1‐AP), P53 antibody (Proteintech, 10442‐1‐AP) and beta‐actin antibody (Proteintech, 7D2C10). The membranes were then incubated with horseradish peroxidase‐conjugated secondary antibodies for 1 hour at room temperature: Goat anti‐rabbit IgG (Proteintech, SA00001‐2) and goat anti‐mouse IgG (Proteintech, SA00001‐1).