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

Fig. 4.

Fig. 4.

UPAT inhibits β-TrCP1– and β-TrCP2–mediated polyubiquitination of UHRF1. (A) Lysates from 293FT cells that had been transfected with the indicated expression constructs and treated with MG132 were subjected to immunoprecipitation with anti-Flag antibody followed by immunoblotting analysis with anti-HA, anti-Flag, or anti-Myc antibody. See also Fig. S5B. (B) Lysates from HCT116 cells transfected with siRNA targeting β-TrCP1 or β-TrCP2 and/or siUPAT were subjected to immunoblotting analysis with anti-UHRF1 or anti–α-tubulin antibody. α-tubulin was used as a loading control. (C) Lysates from HCT116 cells that had been transfected with Myc-tagged β-TrCP2 and/or UPAT were subjected to immunobotting analysis with anti-UHRF1, anti–α-tubulin, or anti-Myc antibody. α-tubulin was used as a loading control. (D) Viability of HCT116 cells transfected with UHRF1 and/or siRNA targeting UPAT was assessed by Cell Titer-Glo assays. Results are expressed as the mean ± SEM (n = 3). *P < 0.05. (E) Lysates from 293FT cells transfected with the indicated expression constructs and treated with MG132 were immunoprecipitated (IP) with anti-Flag antibody followed by immunoblotting analysis with anti-HA, anti-Flag, or anti-Myc antibody. (F) Lysates from 293FT cells that had been transfected with the indicated expression constructs were subjected to immunoblotting analysis with anti-Flag, anti–α-tubulin, or anti-Myc antibody. α-tubulin was used as a loading control. (G) UPAT binds to UHRF1 and interferes with its β-TrCP1– and β-TrCP2–mediated polyubiqutination. UHRF1 stabilized by UPAT up-regulates SCD1 and SPRY4, which are required for the survival of colon cancer cells.