Figure 5

MOF is physically associated with the copia untranslated leader region (ULR) in vivo. Chromatin immunoprecipitation (ChIP) experiments were performed to determine whether MOF is physically associated with copia DNA sequences in vivo. Precipitation reactions performed independently with the MOF, the MSL-1 and MSL-3 antibodies, rabbit IgG antibody (non-specific control) and no antibody were used as templates for quantitative PCR from Kc cells. The average and standard errors for triplicate reactions were plotted to reveal the average fold increase in precipitated copia DNA for each antibody relative to the no antibody reactions. Precipitated DNA was amplified using primers specific for the copia ULR, the roX-1 gene (positive control) and the Spt4 gene (negative control) (see Methods). MOF and other members of the MSL complex have previously been demonstrated to bind to the roX- 1 gene [e.g., 23]. Spt-4 was chosen as a negative control because it is not a target site for the MSL complex [e.g., 22]. The results indicate that MOF, MSL-1 and MSL-3 are all associated with the copia ULR and roX-1 in vivo. There was no binding above background to the Spt-4 gene.