Fig. 4

Analysis of the binding of BmDeaf1 with the CpGI2 in the intragenic promoter of BmCHSA-2b. a Analysis of chromatin immunoprecipitation targets detected by RT-PCR (above) and by qRT-PCR (middle) in the Bm12 cells. The enrichment of the promoter sequence in the immunoprecipitated DNA samples was normalized with DNA presented in the 10% input material. The enriched RT-PCR product of the ChIP assay was sequenced and aligned with − 270 ~ − 300 in CpGI2 (below). b Pull-down experiment with the wild-type or mutant CpGI2 DNA probe and the nuclear proteins from the Bm12 cells that were transfected with BmDeaf1-EGFP vector. The proteins that bound to the CpGI2 probe in the supernatant were visualized by Western blot with the BmDeaf1 antibody. The Bio-met-probe: the probe was labeled with biotin and methyl. c Electrophoretic mobility shift assay (EMSA) of the binding of the nuclear proteins, which were isolated from the Bm12 cells that were transfected with BmDeaf1-EGFP vector, to the CpGI2 probe. The cold probe is the unlabeled CpGI2 probe. The supershifted band was detected by using the BmDeaf1 antibody. d EMSA of the binding of the nuclear proteins isolated from the mid-pupal wings to the CpGI2 probe. The supershifted band was detected by using the BmDeaf1 antibody. IgG was used a negative control for the supershift assay. The sequences of the wild-type, mutant and methylated CpGI2 probe are shown in Additional file 10: Table S1