FIG. 7.
Effects of PADI4 and HDAC1 on pS2 nucleosome occupancy. (A) Schematic description of the FAIRE technique. Infected MDA::ERα cells were placed for 2 days in an E2-free medium and then exposed to 10 nM E2 for 6 h (upper part). Chromatin was then prepared and phenol-chloroform extracted without previous reverse cross-linking (bottom part). (B) MDA::ERα chromatin samples were prepared from cells expressing control RNAi, PADI4 RNAi, HDAC1 RNAi, or PADI4 RNAi + HDAC1 RNAi (RNAi PADI4 + RNAi HDAC1). FAIRE enrichment in pS2 promoter was quantified by real-time PCR on the basis of the threshold cycle value (CT). Calculation details are described in Materials and Methods. The data are presented as means ± the standard errors of four independent experiments. Asterisks indicate the statistical significance as determined by Dunnett post hoc comparisons (**, P < 0.01; ***, P < 0.001).