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. 2016 Jan 14;113(5):1273–1278. doi: 10.1073/pnas.1500992113

Fig. 3.

Fig. 3.

UPAT stabilizes UHRF1 protein by interfering with its ubiquitination and degradation. (A, Upper) qRT-PCR analysis of UHRF1 expression in HCT116 cells transfected with siRNA targeting UPAT. Results are expressed as the mean ± SEM (n = 3). (A, Lower) Cell lysates were subjected to immunoblotting analysis with anti-UHRF1 or anti-Actin antibody. Actin was used as a loading control. (B) HCT116 cells transfected with siRNA targeting UPAT were treated with CHX for the indicated times and then subjected to immunoblotting analysis with anti-UHRF1 or anti–α-tubulin antibody. α-tubulin was used as a loading control. (C) HCT116 cells transfected with siRNA targeting UPAT were cultured in the presence or absence of MG132 and then subjected to immunoblotting analysis with anti-UHRF1 or anti–α-tubulin antibody. α-tubulin was used as a loading control. (D) Lysates from HCT116 cells that had been transfected with siRNA targeting UPAT and treated with MG132 were subjected to immunoprecipitation with anti-UHRF1 antibody followed by immunoblotting analysis with anti-ubiquitin or anti-UHRF1 antibody. (E) Lysates from 293FT cells transfected with HA-tagged β-TrCP2 along with empty vector (Mock) or mutants of UHRF1 were immunoprecipitated (IP) with anti-Flag antibody followed by immunoblotting analysis with anti-HA or anti-Flag antibody.