Fig. 4: TRABID deficiency induces micronuclei through mitosis and autophagy defects.

a, b HeLa or B16F10 cells stably expressing control or TRABID shRNAs were stained with DAPI. Representative images and the percentages of cells with micronuclei are shown. Bar, 10 μm. c FISH analysis of centromeres on HeLa cells stably expressing TRABID shRNA. Boxed areas are enlarged to show micronuclei with centromere signals. Bar, 10 μm. d, e HeLa cells stably expressing control or TRABID shRNAs were transiently transfected with indicated constructs and treated with or without 100 nM bafilomycin A1 for 16 h. Cells were stained with DAPI and then examined for micronuclei by confocal microscopy. Representative images, the percentages of cells with micronuclei, and the expression levels of various proteins are shown. Bar, 10 μm. Blots are representatives of two independent experiments. Notably, GFP-Aurora B was used for transfection in d and its position is marked. f HeLa cells stably expressing control or TRABID shRNAs were transiently transfected with VPS34 and then stained with LC3 antibody and DAPI. Representative images are shown. Boxed areas are enlarged and shown in the inset or below. Bars, 5 μm. Micronuclei that are colocalized or surrounded by LC3 signals are indicated by arrows. Yellow lines indicate the paths along which the relative intensities of LC3 and DAPI were quantified and plotted (labeled with “1” or “2” to indicate the corresponding images). The percentages of cells with LC3⁺ micronuclei are shown. Data in (a), (b), (d), (e), (f) are mean ± SD (n = 3 independent experiments and >20 cells per group per experiment were counted). P values are determined by one-way (a, b, d, e) or two-way (f) ANOVA with Tukey’s post hoc test. Source data are provided as a Source Data file.