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(A) HEK293T cells were transfected with the indicated plasmids and treated with vehicle or MG-132 for 4 hours before harvest. Western blots probed for Pdx1, Flag (Cul3 and Pcif1), and Tubulin (loading control). (B–D) HEK293T cells transfected with the indicated plasmids prior to immunoprecipitation with IgG or anti-Myc–coupled agarose. Western blots probed for HA (Pdx1). Brackets indicate ubiquitinated Pdx1.
