Anti p53

HUVECs were transfected using the Effectence transfection reagent (Qiagen, Suzhou, Jiangsu, China), while HEK293T cells were transfected using the Lipofectamine 2000 Reagent (Invitrogen, Carlsbad, CA, USA). 5-aza (Decitabine), a potent inhibitor of DNA methylation, was from Selleck Chemicals (Shanghai, China). siRNAs were synthesized from Genepharma (Suzhou, China), the sequences of siRNAs are listed in S1 Table. LncRNA Smart Silencer was obtained from RiboBio (Guangzhou, China). Antibodies against KSHV LANA, HMGB2, CMPK1, DNMT1 and Dicer were from Abcam (Cambridge, MA, USA). Anti-Flag was obtained from Cell Signaling Technologies (Beijing, China). Anti-Myc, anti-α-Tubulin, and anti-GAPDH were from Santa Cruz Biotechnology (Dallas, TX, USA). Anti-rabbit immunoglobulin G (IgG), anti-mouse IgG, anti-phosphorylated p53, anti-acetylated p53, anti-p53, and anti-p21 were purchased from Beyotime Institute of Biotechnology (Nantong, Jiangsu, China). Western-blotting analysis was conducted as previously described [81 (link), 82 (link)]. In this study, all Western blotting results were independently repeated at least three times unless otherwise stated.

The ChIP assay was performed as we previously described^{61 (link)}. Briefly, U2OS cells were cross-linked with 1% formaldehyde for 10 min. The ChIP assay was performed by using anti-p53 and the ChIP assay kit (Beyotime Biotechnology, Shanghai, China) according to the manufacturer’s instructions. Normal mouse immunoglobulin G (IgG) was also used as a negative control. The bound DNA fragments were subjected to real-time PCR analysis using the specific primers (Supplementary Table S2).

Supercritical fluid (SCF) extract of Zanthoxylum bungeanum was purchased from Zhengzhou Xuemailong Food Spice Co., Ltd. in Henan Province, China. Both HCT-116 (human colon adenocarcinoma cell line), and HEK293T cells were obtained from the National Collection of Authenticated Cell Cultures in Shanghai, China. McCoy's 5A media, DMEM media, fetal bovine serum (FBS), and 0.25% trypsin solution were obtained from Invitrogen (Carlsbad, CA, USA). The Caspase 8 inhibitor Z-IETD-FMK and P53 inhibitor PFTα hydrobromide were obtained from MedChemExpress (Monmouth Junction, NJ, USA). Primary antibodies (anti-CDK4, anti-cyclin D1, anti-PCNA, anti-BAX, anti-Caspase 3, anti-Cleaved Caspase 3, anti-P53, anti-Bcl-2, anti-Caspase 9, anti-Cleaved Caspase 9, anti-Fas, anti-P21, anti-Caspase 8, anti-Cleaved Caspase 8, and anti-β-actin), anti-mouse, and anti-rabbit secondary were obtained from the Beyotime Institute of Biotechnology (Shanghai, China).