Journal: World Journal of Surgical Oncology

Article Title: Circ_0001741 regulates proliferation and invasion in ESCC via the miR-194-5p/E2F3 axis

doi: 10.1186/s12957-025-04124-2

Figure Lengend Snippet: E2F3 is a miR-194-5p target gene in TE2 and KYSE150 cells A Predicted partially complementary binding sequence between E2F3 mRNA and miR-194-5p B Dual-luciferase reporter assays confirming miR-194-5p binding to the E2F3 3’UTR. C Pull-down assay was used to confirm the interaction of E2F3 and miR-194-5p D Western blot analysis of E2F3 protein levels in TE2 cells after miR-194-5p overexpression. * P < 0.05

Article Snippet: Membranes were incubated overnight at 4 °C with E2F3 primary antibodies (Proteintech, China, Cat No. 27615-1-AP), followed by HRP-conjugated secondary antibodies (Proteintech, China).

Techniques: Binding Assay, Sequencing, Luciferase, Pull Down Assay, Western Blot, Over Expression

Journal: World Journal of Surgical Oncology

Article Title: Circ_0001741 regulates proliferation and invasion in ESCC via the miR-194-5p/E2F3 axis

doi: 10.1186/s12957-025-04124-2

Figure Lengend Snippet: E2F3 overexpression reverses the effects of circ_0001741 knockdown A E2F3 overexpression partially rescues the anti-invasive effects of si-circ_0001741 or miR-194-5p in TE2 and KYSE150 cells B E2F3 overexpression partially restores proliferation suppressed by si-circ_0001741 or miR-194-5p. * P < 0.05

Article Snippet: Membranes were incubated overnight at 4 °C with E2F3 primary antibodies (Proteintech, China, Cat No. 27615-1-AP), followed by HRP-conjugated secondary antibodies (Proteintech, China).

Techniques: Over Expression, Knockdown

Journal: Cell & Bioscience

Article Title: IgD in nucleus of pro-B cells promotes pro-B cells proliferation by regulating E2F3 expression

doi: 10.1186/s13578-025-01490-y

Figure Lengend Snippet: IgD deletion reduces the accessibility of E2f3a gene to inhibit E2f3a expression. A A heatmap representation of ChIP-seq data of transcription factors (TFs) was utilized to evaluate the genomic occupancy of IgD. B Genome browser tracks illustrate the promoter region at the E2f3 target site in pro-B cells, comparing conditions with anti-IgD to those with an isotype IgG control. C EMSA was used to confirm the specific binding of IgD to the E2f3 promoter region, with biotin-labeled sequences of the E2f3 promoter region used as probes. The assay included a non-biotinylated competition probe (Com Probe), representing an unlabeled sequence of the E2f3 promoter, and a mutation probe (Mut Probe) which has identical sequence with targeted nucleotide substitutions in the core E2F binding motif. The reaction system is depicted in the figure. D Supershift assay detects the specific binding of IgD to the E2f3 promoter region, the reaction system is as shown in the figure. E ChIP-PCR was performed to amplify the promoter regions of E2f3a and E2f3b using pro-B cells from WT mice, with genomic DNA from pro-B cells serving as a positive control. F Transcriptional levels of E2F3a and E2F3b in pro-B cell and pre-B cell derived from IgD −/− mice were detected by RT-PCR. G Protein expression levels of E2f3a in pro-B cell from WT and IgD −/− mice were examined by Western blot analysis. Quantification result was presented as grey scale scans with statistical significance denoted by asterisks (**** p < 0.0001)

Article Snippet: Primary antibodies used included anti-mouse β-actin (Zhongshan Golden Bridge, TA-09), anti-mouse Gapdh (Zhongshan Golden Bridge, TA-08), anti-mouse IgD (Santa, sc-53853), anti-mouse IgM (Bethyl, A90-101P), anti-mouse Igκ (SouthernBiotech, 1050-08), and anti-mouse E2f3 (CST, DF12390).

Techniques: Expressing, ChIP-sequencing, Control, Binding Assay, Labeling, Sequencing, Mutagenesis, Positive Control, Derivative Assay, Reverse Transcription Polymerase Chain Reaction, Western Blot