Fig. 1

Chd4 expression and formation of different NURD complexes during gametogenesis. A Expression of chd4 monitored by immunofluorescence in testis sections of 14 dpp and 2 months (adult) old mouse using two distinct antibodies against Chd4, a rabbit polyclonal anti-Chd4 and anti-Plzf (top panel) and mouse monoclonal anti-Chd4 and anti-Sox9 (bottom panel). Arrows indicate examples of Chd4 positive spermatogonia (Plzf) or Sertoli (Sox9) cells. Cells within the punctuated line are pachytene spermatocytes. Similar results were observed independently using three different mice. B Western blot analysis of different NURD components in samples of cells enriched in undifferentiated spermatogonia (Thy1.2+) (Plzf positive), differentiating spermatogonia (c-Kit+) (Stra8 positive) and the flow-through (FT), the latter from the spermatogonia enrichment procedure, which contains mostly somatic cells, including a large amount of Sertoli cells (Sox9 positive). Lamin B (Lam B) and α-tubulin (Tub.) were used as loading standards. The scheme represents the composition of a canonical NURD complex. C Chd4-participating NURD complexes in enriched fractions of spermatogonia or cells in the flow-through assessed by co-immunoprecipitation with anti-Chd4 as a bait. Non-specific IgG was used as a bait control. Experiments were repeated twice, and the star marks an unspecific reactive band