Figure 6

TR binds to the TRE2 in Dot1L promoter in the intestine and tail during T3-induced metamorphosis. Stage 54 tadpoles were treated with or without T3 for 2 days. The intestine (A) or tail (B) was isolated for ChIP assay with a polyclonal antibody against TR (top) or Id14 (bottom), a negative control for antibody specificity. The immunoprecipitated DNA was analyzed by qPCR for the presence of the TRE regions of Dot1L promoter. A region of exon 2 (ex2) of Dot1L gene was analyzed as a negative control for binding specificity. Note that strong binding of TR to TRE2 in Dot1L gene was observed in the absence of T3 in premetamorphic tadpoles and that this binding was increased slightly upon T3 treatment in both organs. TR signal at the TRE1 was very low in both organs with or without T3 treatment, consistent with the results from transcription and in vitro TR binding assays. Only background signals were observed with the anti-ID14 antibody. Error bars indicate s.e.m