(A) TAX1BP1 truncation or point mutations used in this study.
(B) i–x, TAX1BP1 KO and TAX1BP1 KO with stable expression of TAX1BP1 mutants exposed to 5 μg/mL for 2 h were either fixed for imaging or washed and followed for a further 3 h in full media; scale bar represents 10 μm. Larger fields of view in Figure S7A.
(C and D) Quantification of Ub-foci formation (C) or clearance (D) observed in (B).
(E and F) Quantification of Ub-foci formation (E) or clearance (F) observed in TKO (OPTN/NDP52/TAX1BP1) ceils treated as in (B). See Figure S7B for images.
(G) WT cells stably expressing high levels of FLAG-TAX1BP1 exposed to 5 μg/mL puromycin for 2 h were either fixed for imaging or washed and followed for 3 h in full media; scale bar represents 10 μm.
(H and I) Quantification of Ub-foci formation (H) or clearance (I) observed in (G) and in Figure S7C. All images are representative of at least three independent experiments in which percent of cells containing Ub-positive foci was assessed in ~200 cells per condition. All quantification displayed as mean ± SD from three independent experiments using one-way ANOVA test comparing to WT (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001) and Tukey’s post hoc test.