Fig. 2: TRABID is upregulated in M phase to control CPC abundance.

a Experimental workflow of SILAC-based analysis of proteome changes by TRABID knockdown. b Mitotic cell division regulators that are significantly downregulated by TRABID depletion as revealed by LC-MS/MS. Data are mean ± SD, n = 3 independent experiments. P values are determined by two-sided Student’s t-test. c, d RT-qPCR (c) and Western blot (d) analyzes of TRABID mRNA and protein levels in HeLa cells treated with 10 μM lovastatin, 2 μg/ml aphidicolin, 10 μM RO-3306, or 3 μM nocodazole for 18 h. Data are mean ± SD, n = 3 independent experiments. P values are determined by one-way ANOVA with Tukey’s post hoc test. e–g Western blot analysis of indicated proteins in HeLa cells stably expressing control or TRABID shRNAs together with or without TRABID wild type or TRABIDΔN/CS and synchronized in M phase by treatment with 3 μM nocodazole for 18 h. The knockdown efficiencies of shRNAs are shown in (e). h Reciprocal immunoprecipitation analyzes of the interactions between TRABID and CPC components in HeLa cells treated with 3 μM nocodazole for 18 h. i HeLa cells transiently expressing V5-TRABID were synchronized at G2 by treatment with 10 μM RO-3306 for 16 h and then released to proceed the cell cycle for 40 min before monitoring the PLA signal with indicated antibodies. Bar, 10 μm. Blots or images are representatives of three (for d, e, g, h) or two (for f, i) independent experiments. Source data are provided as a Source Data file.