Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: KAP1-Mediated Epigenetic Suppression in Anti-RNA Viral Responses by Direct Targeting RIG-I and MDA5.

doi: 10.4049/jimmunol.2100342

Figure Lengend Snippet: FIGURE 2. KAP1 inhibits RNA viralinduced IFN-b activation. (A) Luciferase analysis of IFN-b reporter activation in HEK293T cells trans- fected with IFN-b reporter plasmid with empty control, TRIM28 (KAP1), TRIM24, TRIM33, or TRIM66 expression plasmid and then infected with SeV for 12 h. (B) Luciferase analysis of IFN-b reporter activation in HEK293T cells transfected with IFN-b reporter plasmid, together with empty control plasmid or KAP1 expression plasmids, and then infected with VSV for 12 h. (C) Western blot analysis of KAP1 expression in mouse PMs from WT or CKO mice. Data shown are means ± SD [n 5 6 in (A), and n 5 5 in (B)]. All experiments were repeated three times. **p < 0.01, ***p < 0.001, by Student t test.

Article Snippet: TRIM33 plasmid (65398) and TRIM66 plasmid (65400) were purchased from Addgene and subcloned into the V5 tag to generate V5-TRIM33 and V5-TRIM66.

Techniques: Activation Assay, Luciferase, Plasmid Preparation, Control, Expressing, Infection, Transfection, Western Blot

Journal: Nature communications

Article Title: Trim33 masks a non-transcriptional function of E2f4 in replication fork progression.

doi: 10.1038/s41467-023-40847-0

Figure Lengend Snippet: 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 com- pared 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. Sig- nificance was determined using Kruskal–Wallis test and Dunn’s multiple compar- isons. 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

Article Snippet: Trim33 expression vectors were a gift of Stefano Piccolo (Addgene plasmids # 20902 and 20903).

Techniques: Western Blot, RNA Sequencing, Knock-Out, Expressing, Plasmid Preparation

Journal: Nature communications

Article Title: Trim33 masks a non-transcriptional function of E2f4 in replication fork progression.

doi: 10.1038/s41467-023-40847-0

Figure Lengend Snippet: Fig. 3 | Deletion of Trim33 promotes E2f4 interactions with chromatin and with the Recql helicase. a Genome browser tracks of E2f4 ChIP-seq in p19/Nras Trim33- WT and Trim33-KO cells; n = 2. b Quantification of ChIP-seq tag coverage at E2f4 peaks in p19/Nras Trim33-WT and Trim33-KO cells. c Relationship between changes (log2FC) in E2f4 binding (normalized E2f4 ChIP-seq tags) and mRNA expression (log2FC) of the encoded gene for p19/Nras Trim33-KO vs Trim33-WT cells. Each dot is a bin of 5 genes sorted by the change in mRNA levels. Trendlines are derived from linear regression analysis. d LC-MS/MS analysis of E2f4 interactome in p19/Nras Trim33-WT and Trim33-KO cells. Known interaction partners are highlighted in blue. e PLA analysis of Recql-E2f4 association in p19/Nras Trim33-WT and Trim33- KO cells stably expressing HA-tagged E2f4 or vector Ctrl; from left, n = 49,49,50,50

Article Snippet: Trim33 expression vectors were a gift of Stefano Piccolo (Addgene plasmids # 20902 and 20903).

Techniques: ChIP-sequencing, Binding Assay, Expressing, Derivative Assay, Liquid Chromatography with Mass Spectroscopy, Stable Transfection, Plasmid Preparation