(A) U2OS cells were co-transfected with pGL3-Hsp27pr-luc, pCruzHA or pCruzHA-LEDGF/p75, and increasing amounts of pcDNA-Flag-MeCP2 (1=1.66 µg DNA). (B) U2OS cells were co-transfected with pGL3-Hsp27pr-luc, pCruzHA or pCruzHA-LEDGF/p52, and increasing amounts of pcDNA-Flag-MeCP2. (C) Transient knockdown of LEDGF/p75 in U2OS cells was achieved using specific siRNA oligos. Cells were then co-transfected with pcDNA-Flag-MeCP2 and pGL3-Hsp27pr-luc. Promoter activity determined as luciferase light units/protein is expressed as fold activation compared to control activity. (D) PC3 cells stably transfected with empty pcDNA vector or pcDNA-LEDGF/p75 were co-transfected with pGL3-Hsp27pr-luc and pMAX-GFP (transfection control). (E) PC3 cells stably transfected with empty pcDNA vector or pcDNA-LEDGF/p75 were co-transfected with pGL3-Hsp27pr-luc and pcDNA-Flag-MeCP2. (F) PC3 cells stably transfected with empty pcDNA vector or pcDNA-LEDGF/p75 were transfected with siRNA oligos to knockdown this protein. Cells were then co-transfected with pGL3-Hsp27pr-luc and pcDNA-Flag-MeCP2. Promoter activity determined as luciferase light units/GFP is expressed as fold activation compared to control activity. Data represent the average of at least three independent experiments. Representative immunoblots corresponding to the reporter assays show protein expression. All protein bands in panel F were from the same blot. *p<0.05; **, p<0.01.