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: International Journal of Molecular Sciences

Article Title: MicroRNA-17-92 Regulates the Transcription Factor E2F3b during Myogenesis In Vitro and In Vivo

doi: 10.3390/ijms18040727

Figure Lengend Snippet: E2F3b is upregulated during myogenesis and is essential for myogenic differentiation.( A ) Western blotting showing increased expression of E2F3a and E2F3b protein in C2C12 myoblasts transfected with vectors expressing E2F3a or E2F3b (OE-E2F3a, OE-E2F3b), compared with the control (empty vector, OE-Ctrl). β-tubulin was used as a loading control; ( B ) Western blotting showing the expression of E2F3a and E2F3b during myogenic differentiation. C2C12 cells were cultured in growth medium (GM) and switched to differentiation medium (DM) for one to seven days. β-tubulin was used as a loading control; ( C ) Western blotting confirmed the efficiency of sh-E2F3 on E2F3a and E2F3b protein expression in C2C12 cells. β-tubulin was used as a loading control; ( D ) Immunostaining of myosin showing that silenced E2F3b resulted in smaller myotubes. C2C12 cells were infected with sh-E2F3 vectors in DM for three days. Scale bar, 200 μm; ( E ) Western blotting showing that silenced E2F3b resulted in a decreased myogenin level at DM3. β-tubulin was used as a loading control. Protein quantitation of myogenin was performed with Image J software. The error bars depict the mean ± SD of three cell samples; ( F ) Immunostaining of myosin showing that OE-E2F3b resulted in longer myotubes. C2C12 cells were lentivirally infected with vectors expressing OE-E2F3 in DM for three days. Scale bar, 200 μm; ( G ) Western blotting showing that OE-E2F3b resulted in increased myogenin protein at DM3. β-tubulin was used as a loading control. Protein quantitation of myogenin was performed with Image J software. The error bars depict the mean ± SD of three cell samples; ( H ) Lentiviral E2F3b rescues the formation of myotubes in sh-E2F3 cells. sh-E2F3 cells were lentivirally infected by E2F3b, transferred to DM, and stained for Myosin heavy chain (MHC) and 4′,6-diamidino-2-phenylindole (DAPI) at DM4; ( I ) Western blotting shows that the introduction of E2F3b increases myogenin protein expression at DM4. Scale bar, 500 μm. The error bars depict the means ± SD of three independent samples. * p < 0.05, *** p < 0.001 and ns, non-significant.

Article Snippet: The following primary antibodies were used: β-actin (1:10,000; Proteintech 66009-1-IG, Wuhan, China), β-tubulin (1:5000; Proteintech 10094-1-AP, Wuhan, China), E2F3 (1:600; Santa Cruz Biotechnology SC-878, Santa Cruz, CA, USA), and myogenin (1:5000; Abcamab124800, Cambridge, MA, USA).

Techniques: Western Blot, Expressing, Transfection, Control, Plasmid Preparation, Cell Culture, Immunostaining, Infection, Protein Quantitation, Software, Staining

Journal: International Journal of Molecular Sciences

Article Title: MicroRNA-17-92 Regulates the Transcription Factor E2F3b during Myogenesis In Vitro and In Vivo

doi: 10.3390/ijms18040727

Figure Lengend Snippet: E2F3b expression is regulated by miR-17-92 in vitro. ( A ) Quantitative real-time PCR (RT-PCR) showing that 56 miRNAs were downregulated in differentiated C2C12myotubes at DM4, compared with myoblasts at GM. The miRNAs computationally predicted to target E2F3b were shown in green; ( B ) Seven miRNAs potentially regulated the expression of E2F3b, including let-7c, miR-15a, miR-17, miR-20a, miR-106a, miR-128, and miR-490; miR-17 and miR-20a, both members of the miR-17-92 cluster were shown in red. ( C ) Quantitative RT-PCR was performed to analyse the expression of miR-17 and miR-20a in C2C12 cells during myogenic differentiation. The data are presented as the means ± SD ( n = 3), the expression in GM was set to 1.0; ( D ) 3′UTR reporter assay showing that miR17/20a and miR-17-92 targeted the 3′UTR of E2F3 in HEK293A cells. The data are presented as the means ± SD ( n = 3). *** p < 0.001; ( E ) miR17/20a and miR-17-92 downregulated the protein expression of E2F3b in differentiated C2C12 cells at DM4. β-actin was used as a loading control. Protein quantitation of E2F3b was performed with Image J software. Values are means ± SD. Data are representative of three independent cell samples. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The following primary antibodies were used: β-actin (1:10,000; Proteintech 66009-1-IG, Wuhan, China), β-tubulin (1:5000; Proteintech 10094-1-AP, Wuhan, China), E2F3 (1:600; Santa Cruz Biotechnology SC-878, Santa Cruz, CA, USA), and myogenin (1:5000; Abcamab124800, Cambridge, MA, USA).

Techniques: Expressing, In Vitro, Real-time Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Reporter Assay, Control, Protein Quantitation, Software

Journal: Biochemical and biophysical research communications

Article Title: A significant role of transcription factors E2F in inflammation and tumorigenesis of nasopharyngeal carcinoma.

doi: 10.1016/j.bbrc.2020.01.158

Figure Lengend Snippet: Fig. 4. Inhibition of E2Fs repressed tumorigenesis in vivo. (A) Experimental procedure of the nude mice subcutaneous tumorigenesis model. (B) Representative photographs of mice tumors corresponding to each treatment group at the last time point. The tumor volume growth curves of the three groups of xenograft tumors were shown in the right panel. (C) Expressions of E2F1 and E2F3 in xenograft tumors were detected by immunohistochemistry. (D) Expressions of CD80 and CD68 in xenograft tumors were detected by immunohistochemistry.

Article Snippet: Antibodies against E2F1 and E2F3 were obtained from Abcam; CD68 and CD80 were obtained from Proteintech (Wuhan, China).

Techniques: Inhibition, In Vivo, Immunohistochemistry

Journal: Molecular Therapy. Nucleic Acids

Article Title: Shrimp miR-34 from Shrimp Stress Response to Virus Infection Suppresses Tumorigenesis of Breast Cancer

doi: 10.1016/j.omtn.2017.10.016

Figure Lengend Snippet: The Mechanism of Shrimp miR-34 in Inhibiting Breast Cancer Progression (A) The prediction of genes targeted by shrimp miR-34. The seed sequence of miR-34 is underlined. (B) The direct interaction between shrimp miR-34 and its target genes (i.e., CCND1 , CDK6 , CCNE2 , E2F3 , MET , or FOSL1 ) in breast cancer cells (MDA-MB-231). Luciferase activity was normalized to the ratio of firefly and Renilla luciferase activities. (C) The effects of shrimp miR-34 expression on the expression of miR-34 target genes in breast cancer cells (MDA-MB-231 and MDA-MB-435). (D) The influence of shrimp miR-34 expression on the expression of miR-34 target gene-encoding proteins in breast cancer cells. Protein levels were detected using western blot analysis. As a control, miR-34-scrambled was included in the transfection. Tubulin was used as a loading control. (E) The loading of shrimp miR-34 onto the human Ago2 complex. Shrimp miR-34 was incubated with the human Ago2 complex, followed by EMSA. Shrimp miR-34 was visualized by ethidium bromide staining (top). Human Ago2 protein was detected with western blot analysis (bottom). The wedge indicates the concentration gradient of human Ago2 complex. (F) A model for the role of miR-34-mediated cancer cell growth and metastasis. In all panels, statistically significant differences between treatments are represented with asterisks (error bar, SD; *p < 0.05 and **p < 0.01).

Article Snippet: Rabbit anti-human CCND1, CDK6, CCNE2, E2F3, FOSL1, and MET antibodies were purchased from Proteintech Group (USA).

Techniques: Sequencing, Luciferase, Activity Assay, Expressing, Western Blot, Control, Transfection, Incubation, Staining, Concentration Assay