Additional file 3: Figure S1. of Mechanisms of Groucho-mediated repression revealed by genome-wide analysis of Groucho binding and activity

Antibody validation (A) Chromatin isolated and sheared exactly as for the ChIP-seq analysis was subjected to immunoprecipitation with the indicated amounts (in μl) of affinity purified antibody against the Gro GP domain used for the ChIP-seq analysis, and then probed in a western blot with both an anti-Gro monoclonal antibody (mAb) or the anti-GP antibody. The band indicated by the asterisk is a cross-reacting protein that is recognized in the western blot but that is not efficiently immunoprecipitated by the anti-GP antibody. Ab HC – antibody heavy chain. (B) Heat map showing overlap (Jacard similarity coefficient [96]) between the peaks called in the duplicate ChIP-seq experiments at each time point. (C) Representative genome browser tracts comparing duplicate ChIP-seq experiments. (D and E) Comparison of Gro binding patterns obtained by ChIP-seq using our anti-GP antibody with that obtained by ChIP-chip (0–12 hr embryos; modENCODE #597) and ChIP-seq (white pre-pupae; modENCODE #4981) using independently derived antibodies [40]. (PDF 588 kb)