Fig. 6

The USP25 dimer stabilizes endogenous tankyrase in HEK293T cells. a Flag-USP25FL, Flag-USP25FL ΔIL, Flag-USP25FL C178A, Flag-USP25FL ΔIL C178A, Flag-USP25NCD, and Flag-USP25NCD ΔIL were transfected in HEK293T cells and the levels of endogenous tankyrases1/2 were analyzed by western blot (WB). GFP was transfected as a control. Plot of the quantification of tankyrase1/2 levels, corrected with tubulin and relative to GFP. Data values are mean ± s.d. and n ≥ 3 technical replicates. Significance was measured by a two-tailed unpaired t test for all lanes relative to GFP and between USP25FL and USP25FL ΔIL. *P < 0.05, **P < 0.01. b HEK293T cells were transfected with Flag-USP25FL, Flag-USP25FL ΔIL, and GFP and cells were treated with 100 μg mL⁻¹ of cyclohexamide (CHX) and collected at indicated times for WB. Endogenous tankyrase1/2, Flag-USP25, and tubulin was checked and compared by WB. c Flag-USP25FL and indicated point mutants were transfected in HEK293T cells and the levels of endogenous tankyrases1/2 were analyzed by WB. Plot of the quantification of tankyrase1/2 levels relative to Flag-USP25FL. Data values are mean ± s.d. and at least n ≥ 3 technical replicates. Significance was measured by a two-tailed unpaired t test relative to FL. *P < 0.05, **P < 0.01. d HEK293T cells were transfected with Flag-USP25FL, Flag-USP25FL P521S, and GFP and cells were treated with 100 μgml⁻¹ of CHX and collected at indicated times for western blotting. Source data are provided as a Source Data file