Fig. 4: SMYD3 regulated S1PR1 via methylation of the S1PR1 promoter.
From: SMYD3 promotes hepatocellular carcinoma progression by methylating S1PR1 promoters
A The specific SMYD3 DNA binding site (–CCCTCC-) in the S1PR1 promoter. B Luciferase assays with the S1PR1 promoter were performed in HCC cells transfected with different amounts of pCDNA-SMYD3 vector in SK-Hep1 cells or cells with stable overexpression (Huh7 cells) or knockdown (Hep3B cells) of SMYD3. C SK-Hep1 cells were cotransfected with pGL-S1PR1-WT, pGL-S1PR1-MT1, pGL-S1PR1-MT2, pGL-S1PR1-MT3, and SMYD3 (pcDNA3.1-SMYD3) (left panel) and Huh7-Ctrl and Huh7-SMYD3 cells were cotransfected with pGL-S1PR1-WT, MT1, MT2 and MT3 (right panel). The promoter activities were measured by performing luciferase reporter assays. D The examined primer position for ChIP-PCR assays at the S1PR1 locus was indicated. E, F The ChIP assay was performed in SK-Hep1-Ctrl and SK-Hep1-SMYD3 cells (E) and Hep3B-sh-NC and Hep3B-sh-SMYD3 (F) cells using specific antibodies. The immunoprecipitated DNA was measured with real-time PCR using the primers listed above. Data are presented as the means ± SD of three separate experiments, each performed in triplicate. *P < 0.05; **P < 0.01.
