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. Author manuscript; available in PMC: 2015 Sep 16.
Published in final edited form as: Nat Commun. 2015 Mar 16;6:6380. doi: 10.1038/ncomms7380

Figure 7. Epsin ENTH-UIM interacted with and impaired polyubiquitination of the Dvl2 C-terminus.

Figure 7

(a) Polyubiquitinated full length or truncated flag-tagged Dvl2 was immunoprecipitated using anti-flag antibody in HEK 293T cells transfected with Flag-tagged Dvl2 full length (Dvl2), DIX-PDZ alone (DIX-PDZ) or DEP-YHEL alone (DEP-YHEL) constructs and HA-tagged ubiquitin and then analyzed by Western blotting. (b) Specified flag-tagged Dvl2 constructs were co-immunoprecipitated using anti-epsin 1 antibody in HEK 293T cells transfected with specified flag-tagged Dvl2 constructs and full length epsin 1 and then analyzed by Western blotting. (c) Epsin 1 was co-immunoprecipitated using anti-flag antibody in HEK 293T cells transfected with Flag-tagged Dvl2 full length (Dvl2), Dvl2 ΔDIX (ΔDIX) or Dvl2 ΔDEP-YHEL (ΔDEP-YHEL) constructs and full length epsin 1 and then analyzed by Western blotting. (d) Polyubiquitinated full length flag-tagged Dvl2 was immunoprecipitated using anti-flag antibody in MG132-treated control (CTL) or epsins 1 and 2 knockdown (KD) HEK 293T cells transfected with flag-tagged Dvl2 and HA-tagged ubiquitin and then analyzed by Western blotting. (e) Polyubiquitinated endogenous Dvl2 was immunoprecipitated using anti-Dvl2 antibody in HEK 293T cells transfected with specified flag-tagged epsin 1 constructs and HA-tagged ubiquitin and then analyzed by Western blotting. (f,g) Dvl2 was co-immunoprecipitated using anti-epsin 1 antibody in HEK 293T cells transfected with specified mutant epsin 1 constructs and Dvl2 and then analyzed by Western blotting. All are representative blots from n=8.