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. Author manuscript; available in PMC: 2019 Dec 6.
Published in final edited form as: Mol Cell. 2018 Nov 8;72(5):823–835.e5. doi: 10.1016/j.molcel.2018.09.019

Figure 5: The E6-Cdt2-USP46 complex is independent of E6AP, p53, UAF1 or USP12.

Figure 5:

(A) Negative control IgG or anti-Cdt2 immunoprecipitates from HeLa cells immunoblotted with indicated antibodies. (B) Same as (A), except with E6AP immunoprecipitates. (C) Same as (A), except with UAF1 (WDR48) immunoprecipitates. (D) Same as (A), except with mycUSP46 immunoprecipitates. (E) HeLa cells transfected with siRNAs indicated at top were immunoblotted as indicated. (F) In vivo ubiquitination of Cdt2 studied by immunoprecipitation of Flag-Cdt2 and immunoblotting, after transfection of HEK-293T cells with plasmids expressing proteins indicated at the top. (G) Lysates from 293T cells transfected with FlagCdt2, Myc-USP46 and Myc-E6 were passed through Superose 6–10/300 column and fractions collected. Even numbered fractions were immunoblotted with indicated antibodies. Peaks of elution of MW markers are indicated at the top. (H) Indicated fractions from gel-filtration chromatography were pooled and immunoprecipitated with either Rabbit IgG or with Cdt2 antibody and probed with Flag or Myc antibodies.