Figure 5.

Knockdown of NIPBL inhibited expression of STAT3-related gene, NIPBL directly regulate the expression of STAT3. (A) H1299 cells were transfected with siNIPBL or non-targeting scramble siRNA for 48 h, RT-PCR analysis of STAT3, c-Myc, and Mcl-1 mRNA levels. * P <0.05. (B) After transfection 72 h, H1299 cells collected and subjected to western blot analysis for detection of NIPBL, STAT3-related proteins and apoptosis-associated proteins levels, a-tubulin was used as a loading control. Shown were representative blots of three independent experiments. (C) Chromatin-immunoprecipitation (ChIP) assays were used to demonstrate that NIPBL bind to STAT3 promoter region. Two pair of primer (STAT3-p1 and STAT3-p2) was designed to amplify different fragments of STAT3 promoter region. Input DNA before immunoprecipitation was used as positive control, as negative control, a nonspecific antibody (a-IgG) was used for precipitation.