Anti dnmt3a sc 365769

The human colorectal cancer cell lines HCT116 and SW480, and mouse colon carcinoma cell line CT26 were cultured in a complete RPMI 1640 growth medium (Thermo Fisher Scientific, CA, USA) with 10% fetal bovine serum (Invitrogen, CA, USA), 3.5 g/L glucose (Thermo Fisher Scientific, CA, USA), 10 mM HEPES (Thermo Fisher Scientific, CA, USA), and 1.0 mM sodium pyruvate (Thermo Fisher Scientific, CA, USA) at 37 °C in an incubator of 5% CO₂ and 95% air.
The following antibodies were used in this study: anti-PD-L1 (ab205921, clone 28-8, Abcam, Cambridge, UK) anti-PD-L1 (#13684, clone E1L3N, Cell Signaling Technology, MA, USA), anti-β-actin (sc-8432, Santa Cruz, CA, USA), anti-DNMT3a (sc-365769, Santa Cruz, CA, USA), anti-DNMT1 (sc-271729, Santa Cruz, CA, USA), anti-p-signal transducer and activator of transcription 1 (p-STAT1, sc-8394, Santa Cruz, CA, USA), anti-STAT1 (sc-464, Santa Cruz, CA, USA), anti-interferon regulatory factor 1 (IRF1, sc-514544, Santa Cruz, CA, USA), and horseradish peroxidase (HRP)-conjugated secondary antibodies (Santa Cruz, CA, USA).
Lentiviruses carrying individual shRNA were obtained from the National Core Facility for Manipulation of Gene Function by RNAi, miRNA, miRNA sponges, and CRISPR/Genomic Research Center, Academia Sinica, Taipei, Taiwan.