Fig. 8.

Identification of STAT3‐binding sites in MLST8 promoter. (A) Protein–DNA complexes from HCT116 cells were precipitated with either STAT3 antibody or normal IgG and the amount of DNA was determined by PCR with primers for promoter regions of the MLST8 gene, followed by agarose electrophoresis (left) or quantitative PCR (right; n = 3). (B) Luciferase reporter assay was performed in HCT116 cells treated with 5 nm of siCTRL or siSTAT3. MLST8 promoter‐luciferase constructs and pCMV3.1‐Renilla vector were transfected 48 h later, and the cells were further incubated for 24 h before harvest. Firefly luciferase activity was normalized with Renilla luciferase activity (n = 4). Data are presented as mean ± SEM. Statistically significant differences are marked with *P < 0.05, **P < 0.01, and ***P < 0.001, respectively; ns, statistically insignificant (A, t‐test; B, two‐way ANOVA).