Journal: Oncogene

Article Title: PMM2 interacts with TRIM28 to recruit E2F4 and promote KIFC3-mediated tumor glycolysis and colorectal cancer progression

doi: 10.1038/s41388-026-03707-x

Figure Lengend Snippet: A Immunoprecipitation combined with mass spectrometry (IP-MS) analysis of TRIM28-interacting proteins in HCT-116 cells. Co-immunoprecipitation (Co-IP) assays in HCT-116 cells validating the interaction between endogenous TRIM28 and E2F4 ( B : IP with anti-TRIM28; C : IP with anti-E2F4). RT-qPCR ( D ) and Western blot ( E ) analysis of KIFC3 mRNA and protein levels in HCT-116 cells with E2F4 overexpression. F Dual-luciferase reporter assay in 293 T cells transfected with WT or Mut1 KIFC3 promoter constructs, with or without E2F4 overexpression. G ChIP assay showing E2F4 enrichment at the KIFC3 promoter site 1 in HCT-116 cells. H Dual-luciferase reporter assay in 293 T cells co-transfected with PMM2, E2F4, and KIFC3 promoter constructs. I ChIP assay showing PMM2 overexpression enhances E2F4 binding to the KIFC3 promoter site 1 in HCT-116 cells. RT-qPCR ( J ) and Western blot ( K ) analysis of KIFC3 expression in HCT-116 cells with PMM2 overexpression and/or TRIM28 knockdown. L Dual-luciferase reporter assay in 293T cells co-transfected with PMM2, TRIM28 shRNA, E2F4 shRNA, and KIFC3 promoter constructs. M ChIP assay showing TRIM28 knockdown partially inhibits PMM2-induced E2F4 binding to the KIFC3 promoter site 1 in HCT-116 cells. Data are mean ± SD ( n = 3). * p < 0.05, ** p < 0.01 vs. corresponding control. ns, not significant.

Article Snippet: Anti-PMM2 (10666-1-AP), anti-E2F4 (10923-1-AP), anti-GAPDH (60004-1-lg), anti-β-actin (66009-1-lg), anti-LDHA (19987-1-AP), anti-PKM2 (15822-1-AP), anti-TRIM28 (15202-1-AP), anti-Histone H3 (17168-1-AP), anti-Flag (66008-3-lg) and anti-HA (51064-2-AP) were purchased from Proteintech; Goat anti-mouse (A0216) and goat anti-rabbit (A0208) were obtained from Beyotime.

Techniques: Immunoprecipitation, Mass Spectrometry, Protein-Protein interactions, Co-Immunoprecipitation Assay, Quantitative RT-PCR, Western Blot, Over Expression, Luciferase, Reporter Assay, Transfection, Construct, Binding Assay, Expressing, Knockdown, shRNA, Control

Journal: PLoS ONE

Article Title: Establishment of the TBX-code reveals aberrantly activated T-box gene TBX3 in Hodgkin lymphoma

doi: 10.1371/journal.pone.0259674

Figure Lengend Snippet: (A) Potential TF binding sites at the TBX3 locus obtained from the UCSC genome browser (hg19). Binding sites for the TFs STAT5, GATA and E2F are highlighted. (B) RQ-PCR analysis of TBX3 after siRNA-mediated knockdown of STAT5A, STAT5B, GATA3, and E2F4. The data indicated that E2F4 activated TBX3 while STAT5A, STAT5B and GATA3 showed no transcriptional impact. (C) RNA-seq data show expression levels of E2F4 in 100 leukemia/lymphoma cell lines using dataset E-MTAB-7721. Western blot analysis of E2F4 protein in HL cell lines HDLM-2, KM-H2 and L-428, demonstrating high levels in KM-H2. TUBA served as loading control (insert). (D) RQ-PCR analysis of E2F4 expression in HL cell lines, confirming elevated transcript levels in KM-H2. (E) Genomic profiling data for chromosome 16 of HL cell line KM-H2, demonstrating amplification of E2F4 at 16q22. (F) ChIP data from ENCODE for E2F4 in HELA-S3 cells demonstrating binding at the TBX3 promoter.

Article Snippet: The following antibodies were used: alpha-Tubulin (Sigma, #T6199), TBX3 (Thermo Fisher, #711087), E2F4 (Cell Signaling Technology, #40291), BCL2L1 (Cell Signaling, #2764), and CDKN2A (Abcam, #ab108349).

Techniques: Binding Assay, Knockdown, RNA Sequencing, Expressing, Western Blot, Control, Amplification

Journal: PLoS ONE

Article Title: Establishment of the TBX-code reveals aberrantly activated T-box gene TBX3 in Hodgkin lymphoma

doi: 10.1371/journal.pone.0259674

Figure Lengend Snippet: This diagram summarizes the results of this study, placing TBX3 at the centre of an aberrant gene regulatory network. TBX3 is directly targeted by genomic amplification just like its activators E2F4, KDM2B and FOXC1. TBX3 inhibits expression of CDKN2A, NFKBIB and CD19, thereby impacting proliferation, NFkB-signaling and B-cell differentiation, respectively.

Article Snippet: The following antibodies were used: alpha-Tubulin (Sigma, #T6199), TBX3 (Thermo Fisher, #711087), E2F4 (Cell Signaling Technology, #40291), BCL2L1 (Cell Signaling, #2764), and CDKN2A (Abcam, #ab108349).

Techniques: Amplification, Expressing, Cell Differentiation