The mechanisms by which the BCL6/BCoR complex blocks Notch-dependent transcription.
A: HA-tagged Su(H), Flag-tagged MAM1 or/and BCL6 was expressed in embryos and protein extracts were isolated from 50 embryos at stage 10. Co-immunoprecipitation with α-Notch antibody was performed. B: Co-immunoprecipitation using in vitro synthesized proteins. C, D: Flag-tagged MAM1, Flag-tagged dn-MAM1, BCL6 or/and Myc-tagged NBD-S was expressed in embryos and protein extracts were isolated from 50 embryos at stage 10 for each experiment. E, F: Myc-tagged BCoR was expressed in embryos without (E) or with (F) the BCL6 MO, and protein extracts were isolated from 50 embryos at stage 10. Co-immunoprecipitation with α-Notch antibody was performed. α-vimentin antibody was used for a mock immunoprecipitation. G: The expression of Pitx2 at stage 25 was tested by whole mount in situ hybridization and the ratios of Pitx2-expressing embryo number versus total tested embryo number are shown. Total numbers of each injection are shown as “n” on the top of each bar.