Fig. 1: Deletion of Trim33 abates replicative stress.

a Immunoblotting analysis of p19/Nras Trim33-WT and Trim33-KO cell lines; n = 3. b Analysis of the RNA-seq data for two Trim33-KO cell lines; n = 3. Venn diagram shows the number of commonly deregulated genes (p adj <0.01, any log2FC) for the two knockout cell lines compared to Trim33-WT cells. Top three enriched KEGG pathways are shown (analyzed by Enrichr⁷⁰). c DNA fiber assays in Trim33-WT and Trim33-KO cells expressing Myc or a Ctrl vector; n = 2. 100 fibers were measured per sample; scale bar = 1 μm. Significance was determined using Kruskal–Wallis test and Dunn’s multiple comparisons. d Immunoblotting analysis in Trim33-WT and Trim33-KO cells expressing Myc or Ctrl vector; n = 3. e DNA fiber assays in Trim33-WT and Trim33-KO cells, unchallenged (Ctrl) or released from a 4-h hydroxyurea (HU) treatment (HU-rel); n = 3. 100 fibers were measured per sample; significance was determined as in c). Scale bar = 1 μm. f EdU incorporation analysis in Ctrl and HU-treated Trim33-WT and Trim33-KO cells; n = 2. >240 cells per group were quantified; scale bar = 50 µm. Significance was determined as in c). g Immunoblotting analysis in Trim33-WT and Trim33-KO cells at indicated timepoints following release from HU; n = 3. h FACS analysis of PI-stained Trim33-WT and Trim33-KO cells treated with HU for 12 h, and after a 12 h release from the treatment. The graph shows an average of three technical replicates. Significance was determined using one-way ANOVA and Tukey’s multiple comparisons. i Analysis of EdU incorporation during release from synchronization in G1 by serum deprivation; n = 1. At least 416 cells were analyzed per sample; the data were analyzed using Kruskal–Wallis test with Dunn’s multiple comparisons. c, e, f, i Boxplots represent median±quartiles with whiskers ranging up to 1.5-fold of the interquartile range. Source data are provided as a Source Data file.