The CCN₉GG motifs contribute to Bcl-2 promoter activation by RTA.

<p>(A) Scheme showing pGL3-ΔRRE1,2 and pGL3-ΔP1 construct used in this study. (B) Fix amounts (10 µg) of indicated mutant Bcl-2 promoter luciferase constructs were transfected or cotransfected into ten million HEK 293 cells with 10, 20 µg of pcDNA-RTA respectively. Cell lysates were prepared and the luciferase assay was carried out as above mentioned in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0023892#pone-0023892-g004" target="_blank">Fig. 4</a> legend. Deletion of the first and second CCN₉GG motifs led to a significant decrease in RTA-mediated Bcl-2 promoter activity. (C) A ChIP assay was carried out using induced or uninduced BCBL1 cells. RTA-DNA complexes were specifically precipitated with or without RTA antibody. The precipitated DNA was recovered by PCR with primers covering RRE 1 and 2 (upper panel). Real-time PCR was performed to quantify the immunoprecipitated promoter DNA.