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2 protocols using cmpk1

1

Quantifying miR-130b and CMPK1 in Gastric Cancer

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Expression levels of miR-130b and CMPK1 mRNA in gastric cancer cell lines (MGC-803, AGS) were detected by qRT-PCR analysis and summarized in Supplementary Methods. Expression levels of CMPK1 (Abcam, US) and β-actin (Santa Cruz Biotechnology, US) proteins in GC cell lines were detected by western blot analysis. The brief description of western blot was also summarized in Supplementary Methods.
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2

Molecular Mechanisms of Epigenetic Regulation

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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.
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