FIG. 5.

Rgt1 is synergistically recruited to multiple binding sites in vivo. (A) The same plasmids used for the repression assay shown in Fig. 4 were used for a ChIP assay. The arrows indicate the locations and orientations of Rgt1 binding sites. (B) Chromatins prepared from cells (YM4127) containing the reporter plasmids grown under repressing conditions were immunoprecipitated with anti-Rgt1 antibody. Rgt1 binding sites in the immunoprecipitated DNA (IP) were PCR amplified using a primer set (OM3128 and OM3129) containing [α-³²P]dATP and analyzed in a 6% polyacrylamide gel. (C) The amounts of the input DNA and immunoprecipitated DNA were measured by a PhosphorImager, quantified by the ImageQuaNT program, and presented as the ratio of immunoprecipitated counts to input counts. Fig. 8C, box, shows the control of ChIP using anti-Rgt1 antibody.