Fig. 4: OTULIN Tyr56 phosphorylation upon DNA damage promotes its association with β-catenin.

a Co-IP assay with anti-OTULIN antibody in MDA-MB-231 cells treated by Dox (2 µg/ml) and MG132 (10 µM) for 8 h. b Co-IP assay with anti-β-Catenin antibody in MDA-MB-231 cells treated as in a. c Mapping of β-Catenin domain required for interacting with OTULIN. d Mapping of OTULIN domain required for interacting with β-Catenin. e MDA-MB-231 cells transfected with OTULIN WT or Y56D mutant were treated with MG132 (10 µM) for 8 h, then with MG132 in combination of Dox (2 µg/ml) for 8 h. f The Co-IP analysis of Flag-β-Catenin and HA-OTULIN WT, Y56F or Y56D in HEK293T cells. MG132 (10 µM) was added to the culture 16 h before harvest. g Detection of OTULIN phosphorylation at Tyr56 in MDA-MB-231 cells treated with Dox (2 µg/ml) for times indicated. h Linear ubiquitination of precipitated β-Catenin from MDA-MB-231 cells treated with Dox (2 µg/ml) for times as indicated. i TOPFlash assay in MDA-MB-231 cells transfected with OTULIN WT, Y56F, or Y56D. n = 3 independent experiments. p1 = 0.000300, p2 = 0.000286, p3 = 0.00247. The statistical analysis in i was performed by two-sided unpaired t-test and p value is indicated as ^∗p < 0.05, ^∗∗p < 0.01, and ^∗∗∗p < 0.001. Data are presented as Mean ± SD. Source data are provided as a Source Data file.