primary antibody e2f3  (Santa Cruz Biotechnology)

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: Aging (Albany NY)

Article Title: Brevilin A shows an anti-tumor role in prostate cancer via the lncRNA H19/miR-194/E2F3 signaling pathway

doi: 10.18632/aging.204744

Figure Lengend Snippet: The impact of Brevilin A on lncRNA H19, miR-194, and E2F3 expressions. DU145 cells were treated at the concentrations of 10 μM and 20 μM, which were calculated through the IC 50 values of Brevilin A. ( A – I ) qRT-PCR determined lncRNA H19 ( A – C ), miR-194 ( D – F ) and E2F3 mRNA ( G – I ) expressions. ( J – L ) Western blot measured E2F3 expression. * P < 0.05, ** P < 0.01, *** P < 0.001 (vs. control), n = 3.

Article Snippet: Primary anti-E2F3 antibody (1:100) (Proteintech, Wuhan, China) or Ki67 antibody (1:100) (Abcam, ab15580, MA, USA) was added for maintaining at 4°C overnight.

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Control

Journal: Aging (Albany NY)

Article Title: Brevilin A shows an anti-tumor role in prostate cancer via the lncRNA H19/miR-194/E2F3 signaling pathway

doi: 10.18632/aging.204744

Figure Lengend Snippet: The profiles of lncRNA H19, miR-194, and E2F3 in prostate cancer tissues and cell lines. ( A , B ) qRT-PCR detected lncRNA H19 expression in prostate cancer tissues and cell lines. ( C , D ) qRT-PCR determined miR-194 expression in prostate cancer tissues and cell lines. ( E , F ) qRT-PCR examined E2F3 mRNA expression in prostate cancer tissues and cell lines. ( G , H ) Western blot was performed for assaying E2F3 protein level. ( I ) Immunohistochemistry checked E2F3 expression in prostate cancer tissues. Scale bar = 50 μm. * P < 0.05, ** P < 0.01, *** P < 0.001 (vs. Normal or PrEC), n = 3.

Article Snippet: Primary anti-E2F3 antibody (1:100) (Proteintech, Wuhan, China) or Ki67 antibody (1:100) (Abcam, ab15580, MA, USA) was added for maintaining at 4°C overnight.

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Immunohistochemistry

Journal: Aging (Albany NY)

Article Title: Brevilin A shows an anti-tumor role in prostate cancer via the lncRNA H19/miR-194/E2F3 signaling pathway

doi: 10.18632/aging.204744

Figure Lengend Snippet: The interaction among lncRNA H19, miR-194, and E2F3. ( A , B ) Starbase database the binding site between lncRNA H19 and miR-194 as well as the binding site between miR-194 and E2F3. ( C – F ) qRT-PCR examined the profiles of lncRNA H19 and miR-194. ( G , H ) Dual luciferase reporter gene assay measured luciferase activity of DU145 cells. ( I , J ) The miR-194 level was probed via qRT-PCR. ( K – R ) Western blot and qRT-PCR tested the effects of lncRNA H19 and miR-194 on E2F3 expression. ns P > 0.05, *** P < 0.001, n = 3.

Article Snippet: Primary anti-E2F3 antibody (1:100) (Proteintech, Wuhan, China) or Ki67 antibody (1:100) (Abcam, ab15580, MA, USA) was added for maintaining at 4°C overnight.

Techniques: Binding Assay, Quantitative RT-PCR, Luciferase, Reporter Gene Assay, Activity Assay, Western Blot, Expressing

Journal: Aging (Albany NY)

Article Title: Brevilin A shows an anti-tumor role in prostate cancer via the lncRNA H19/miR-194/E2F3 signaling pathway

doi: 10.18632/aging.204744

Figure Lengend Snippet: The influence of lncRNA H19 and miR-194 on DU145 cell proliferation, apoptosis, invasion, and migration. H19 overexpression plasmids and si-H19 were transfected into DU145 cells, and miR-194 mimics or inhibitors were administered for 24 hours culture. ( A , B ) CCK8 assay detected cell proliferation. ( C , D ) Transwell monitored cell migration and invasion. Scale bar = 100 μm. ( E , F ) TUNEL staining examined apoptosis. Scale bar = 50 μm. ( G , H ) qRT-PCR examined the profiles of lncRNA H19 and miR-194. ( I , J ) Western blot was used for examining E2F3 protein level. ** P < 0.01, *** P < 0.001 (vs. NC or siNC); && P < 0.01, &&& P < 0.001 (vs. H19 or siH19), n = 3.

Article Snippet: Primary anti-E2F3 antibody (1:100) (Proteintech, Wuhan, China) or Ki67 antibody (1:100) (Abcam, ab15580, MA, USA) was added for maintaining at 4°C overnight.

Techniques: Migration, Over Expression, Transfection, CCK-8 Assay, TUNEL Assay, Staining, Quantitative RT-PCR, Western Blot

Journal: Aging (Albany NY)

Article Title: Brevilin A shows an anti-tumor role in prostate cancer via the lncRNA H19/miR-194/E2F3 signaling pathway

doi: 10.18632/aging.204744

Figure Lengend Snippet: The impact of miR-194 and E2F3 on DU145 cell proliferation, apoptosis, invasion, and migration. E2F3 overexpression and si-E2F3 plasmids were transfected into DU145 cells for 24 hour culture after miR-194 mimics or inhibitors were administered. ( A , B ) CCK8 assay checked cell proliferation. ( C , D ) Transwell assay examined cell migration and invasion. Scale bar = 100 μm. ( E , F ) TUNEL staining detected cell apoptosis. Scale bar = 50 μm. ( G , H ) qRT-PCR examined the profiles of lncRNA H19 and miR-194. ( I , J ) Western blot was used for examining E2F3 protein level. *** P < 0.001 (vs. NC or NC-in); & P < 0.05, && P < 0.01, &&& P < 0.001 (vs. miR-149 or miR-149 in), n = 3.

Article Snippet: Primary anti-E2F3 antibody (1:100) (Proteintech, Wuhan, China) or Ki67 antibody (1:100) (Abcam, ab15580, MA, USA) was added for maintaining at 4°C overnight.

Techniques: Migration, Over Expression, Transfection, CCK-8 Assay, Transwell Assay, TUNEL Assay, Staining, Quantitative RT-PCR, Western Blot

Journal: Aging (Albany NY)

Article Title: Brevilin A shows an anti-tumor role in prostate cancer via the lncRNA H19/miR-194/E2F3 signaling pathway

doi: 10.18632/aging.204744

Figure Lengend Snippet: Brevilin A hampered the pro-cancer function of lncRNA H19. Brevilin A was utilized to treat DU145 cells stably transfected with lncRNA H19 overexpression plasmids. ( A ) CCK8 assay checked cell proliferation. ( B ) Transwell assay examined cell migration and invasion. Scale bar = 100 μm. ( C ) TUNEL staining detected cell apoptosis. Scale bar = 50 μm. ( D , E ) qRT-PCR examined the profiles of lncRNA H19 and miR-194. ( F ) Western blot was used for examining E2F3 protein level. ** P < 0.01, *** P < 0.001 (vs. NC); & P < 0.05, && P < 0.01, &&& P < 0.001 (vs. H19), n = 3.

Article Snippet: Primary anti-E2F3 antibody (1:100) (Proteintech, Wuhan, China) or Ki67 antibody (1:100) (Abcam, ab15580, MA, USA) was added for maintaining at 4°C overnight.

Techniques: Stable Transfection, Transfection, Over Expression, CCK-8 Assay, Transwell Assay, Migration, TUNEL Assay, Staining, Quantitative RT-PCR, Western Blot

Journal: Aging (Albany NY)

Article Title: Brevilin A shows an anti-tumor role in prostate cancer via the lncRNA H19/miR-194/E2F3 signaling pathway

doi: 10.18632/aging.204744

Figure Lengend Snippet: The functions of Brevilin A and lncRNA H19 in the prostate cancer nude mouse xenograft model. DU145 cells, stably transfected with lncRNA H19 overexpression plasmids, were taken to construct a nude mouse xenograft model of prostate cancer, and Brevilin A was harnessed for treatment. The tumors were excised 28 days later. ( A ) The tumor volume. ( B ) The images of tumors. ( C ) The tumor masses. ( D , E ) Immunohistochemistry determined Ki67 and E2F3 expressions. ( F , G ) qRT-PCR examined the profiles of lncRNA H19 and miR-194. ( H ) Western blot was used for examining E2F3 protein level. *** P < 0.001 (vs. NC); && P < 0.01, &&& P < 0.001 (vs. H19), n = 5.

Article Snippet: Primary anti-E2F3 antibody (1:100) (Proteintech, Wuhan, China) or Ki67 antibody (1:100) (Abcam, ab15580, MA, USA) was added for maintaining at 4°C overnight.

Techniques: Stable Transfection, Transfection, Over Expression, Construct, Immunohistochemistry, Quantitative RT-PCR, Western Blot

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: The paraffin sections were then incubated with primary antibodies against human CCND1, CDK6, CCNE2, E2F3, or MET (Proteintech Group, USA) at 4°C overnight, followed by incubation with HRP-conjugated goat anti-Rabbit IgG (Sigma-Aldrich, USA) for 2 hr at room temperature.

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

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: Molecular Medicine Reports

Article Title: miR-34a suppresses proliferation and induces apoptosis of human lens epithelial cells by targeting E2F3

doi: 10.3892/mmr.2016.5901

Figure Lengend Snippet: E2F3 was a direct target of miR-34a. (A and B) Schematic representation of WT and mut putative miR-34a-binding sites in the 3′-UTR of E2F3. (C) The 293T cells were transfected with miR-34a mimic and psiCHECK2 containing either the WT or mut putative binding sites for miR-34a, and 48 h later the luciferase assay was performed. The data indicated that miR-34a mimic reduced the Renilla activity in the reporter construct containing the E2F3 site (1.87±0.13), whereas no effect was observed with a construct containing a mut E2F3 seed site (3.46±0.09) and mimic control (3.27±0.04). Data are presented as the mean ± standard deviation; *P<0.05. miR, microRNA; WT, wild-type; mut, mutant; UTR, untranslated region; NS, non-significant.

Article Snippet: Subsequent to blocking in Tris-buffered saline with Tween-20 (TBST) containing 25 mmol/l Tris-HCl, pH 7.5, 137 mmol/l NaCl, 2.7 mmol/l KCl and 0.05% Tween-20 with 5% nonfat milk for 1 h at 37°C, the membranes were incubated with the primary antibody against E2F3 (sc-878; 1:200; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) or GAPDH (ab9485; 1:2,500; Abcam, Cambridge, MA, USA) in TBST with 5% nonfat milk at 4°C overnight.

Techniques: Binding Assay, Transfection, Luciferase, Activity Assay, Construct, Control, Standard Deviation, Mutagenesis

Journal: Molecular Medicine Reports

Article Title: miR-34a suppresses proliferation and induces apoptosis of human lens epithelial cells by targeting E2F3

doi: 10.3892/mmr.2016.5901

Figure Lengend Snippet: miR-34a and siE2F3 reduce E2F3 expression. SRA01/04 cells transfected with miR-34a mimic or siE2F3 exhibited reduced E2F3 expression compared with the mock and mimic control. Data are presented as the mean ± standard deviation; *P<0.05. miR, microRNA; si, small interfering; NS, non-significant.

Article Snippet: Subsequent to blocking in Tris-buffered saline with Tween-20 (TBST) containing 25 mmol/l Tris-HCl, pH 7.5, 137 mmol/l NaCl, 2.7 mmol/l KCl and 0.05% Tween-20 with 5% nonfat milk for 1 h at 37°C, the membranes were incubated with the primary antibody against E2F3 (sc-878; 1:200; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) or GAPDH (ab9485; 1:2,500; Abcam, Cambridge, MA, USA) in TBST with 5% nonfat milk at 4°C overnight.

Techniques: Expressing, Transfection, Control, Standard Deviation