Fig. 5

Analysis of the binding of Bmara with the − 250 to − 225 nt fragment of BmCHT10 promoter. a Electrophoretic mobility shift assay (EMSA) of the binding of the purified Bmara proteins to the biotin-labeled probe of wild type − 250 to − 221 nt fragment. The cold probe is the unlabeled probe. b DNA pull-down experiment with the wild type or mutated − 250 to − 225 nt fragment and the nuclear proteins from the Bm12 cells that were transfected with Bmara-FLAG overexpression vector. The proteins that bound to the − 250 to − 225 nt probe in the supernatant were visualized by Western blot with the antibody against FLAG. c qRT-PCR and Western blot analyses showed that the mRNA and protein levels of Bmara in the Bm12 cells were increased when Bmara-FLAG was overexpressed. d The chromatin immunoprecipitation targets were detected by RT-PCR and qRT-PCR (left) in the Bm12 cells. The enriched RT-PCR product of the ChIP assay was sequenced and the sequence was aligned with the − 250 to − 225 nt fragment (right). Each data point is the mean ± SE of three independent assays. For the t test: p < 0.05 (*) or p < 0.01(**)