Figure 6.

RoX2 affects the DNA binding of MSL2^vir in different ways. (A) Venn diagram showing overlapping peaks of three independent MSL3 ChIP experiments using female melanogaster cells transfected with the MSL2^vir-GFP (VIR) construct with (+roX2) or without the roX2 gene and MSL2^mel in vivo binding sites (HASs). Only peaks common to all replicates are considered. (B) Western blot analyses of pull-down experiments using the recombinant MSL2^mel-FLAG (MEL) or MSL2^vir-FLAG (VIR) constructs as bait to retrieve interacting proteins from extracts of male melanogaster cells, untreated or treated with RNase A. (C) Representative DIP-seq profiles of MSL2^vir (VIR) in the presence of no RNA (VIR), of a control RNA (CTRL), or of the roX box RNA (RB4). (D) Read counts on 9534 ensemble binding sites from DIP-seq experiments of MSL2^vir in the presence of no RNA (VIR), of control RNA (CTRL), or of the roX box RNA (RB4). Two independent experiments are shown. (E) FRAP analyses of female melanogaster cells expressing either MSL2^vir-GFP (nucleoplasm) or MSL2^vir-GFP + roX2 (X). Relative fluorescence intensity measured in the bleached area was plotted against recovery time. Shade curves represent the standard error of the measurements. N = 9.