Research Paper Volume 12, Issue 8 pp 7282—7298

HMGN5 promotes IL-6-induced epithelial-mesenchymal transition of bladder cancer by interacting with Hsp27

Figure 6. HMGN5/Hsp27 modulates IL-6-induced STAT3 phosphorylation and Twist promoter activity. (A) J82 and T24 cells were transduced with Lv-sh-HMGN5 under IL-6 stimulation and examined for the protein levels of p-STAT3, STAT3, p-Hsp27, and Hsp27. (B) ChIP assays were conducted on nuclear extracts from T24 cells cotransduced with Lv-sh-HMGN5 and Lv-Hsp27 and treated with or without IL-6, after which immunoprecipitations were conducted with anti-IgG or anti-STAT3 and Protein G agarose. Real-time PCR was conducted using immunoprecipitated DNAs, soluble chromatin, and specific primer pairs for Twist. The results of immunoprecipitated samples were corrected for the results of the corresponding soluble chromatin samples. (C) T24 cells were treated as described above and were cotransfected with 0.5 μg/mL of Twist-Luc plasmid and 0.05 μg/mL pRL-TK and treated with or without 50 ng/mL IL-6 for 6 h before lysis. The luciferase activity of Twist-Luc–969 alone was set as 1. The data are presented as the mean ± SD of three independent experiments. **P<0.01.