Fig. 1. p62, NBR1, and TAX1BP1 colocalize in ubiquitin-containing condensates.

a The cargo receptors NBR1 and p62 were endogenously tagged with mScarlet-AID and GFP tags respectively, using CRISPR (Supplementary Fig. 1b). Cells were left untreated (DMSO) or treated with bafilomycin (400 nM for 2 h) and fixed. After fixation, NBR1 and p62 were detected using their endogenous fluorescent tags, while TAX1BP1 was detected by immunofluorescent staining. Scale bar, 10 µm. Validation of endogenous protein tagging in the cell line used for the experiment is shown in Supplementary Fig. 1c and d. b, c Colocalization of TAX1BP1 with p62, NBR1 or both (b) and colocalization of p62 with TAX1BP1, NBR1 or both (c), based on the experiments in Fig. 1a. Colocalization analysis was performed with ImageJ. Average percentage of colocalization and SEM for three independent experiments are plotted. An unpaired, two-tailed Student’s t test was used to estimate significance. P values are indicated in the figure. d Colocalization of TAX1BP1 with LC3B in HAP1 WT cells mock-treated with DMSO or treated with bafilomycin (400 nM) for 2 h. LC3B and TAX1BP1 were detected by immunofluorescence staining. Scale bar, 10 µm. For the colocalization analysis, average percentages of colocalization ± SEM for n = 3 are plotted. An unpaired, two-tailed Student’s t test was used to estimate significance. P values are indicated in the figure.