e2f4 (a-20) rabbit polyclonal antibody  (Santa Cruz Biotechnology)

Journal: International Journal of Biological Sciences

Article Title: Transcription Factor E2F4 Promote Proliferation, Migration, and Invasion of Gastric Cancer Cells by transcriptionally activating DSCC1

doi: 10.7150/ijbs.99590

Figure Lengend Snippet: E2F4 Regulation of DSCC1 Transcription and Expression in GC. (A) Detection of differentially expressed genes (DEGs) within the major cluster as targets identified by enriched motifs/tracks associated with the E2F4 transcription factor. (B) Heatmap illustrating the expression of transcription factors regulating DSCC1 in GC and normal samples. (C) Analysis of expression correlations between E2F4 and PLAUR based on TCGA-seq data. (D) Representative confocal microscopy image demonstrating the co-localization of E2F4 protein (green) and DSCC1 (red) in AGS and NCI-N87 cell lines. (E-F) Positive regulatory effect of E2F4 on DSCC1 expression in GC cells. Evaluation of E2F4 silencing on DSCC1 mRNA and protein expression in GC cells using RT-qPCR (E) and Western blot (F) , respectively. (G) Prediction of the E2F4 binding motif within the DSCC1 promoter through the JASPAR dataset. (H) ChIP-qPCR analysis indicating E2F4 binding to the DSCC1 promoter in AGS and NCI-N87 cell lines, with the first position being the most significant binding site. (I) Luciferase activity assessment following mutation of the first E2F4 site in the DSCC1 promoter in 293T cell lines. The luciferase activity of the wild-type DSCC1 promoter was notably higher than that of the mutant DSCC1 promoter. (*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001) - Significance levels indicated by asterisks.

Article Snippet: A portion of the lysates was designated as “Input.” The remaining lysates were incubated with E2F4 antibody (CST, #40291) and Protein G magnetic beads to form a DNA-antibody-magnetic bead complex.

Techniques: Expressing, Confocal Microscopy, Quantitative RT-PCR, Western Blot, Binding Assay, ChIP-qPCR, Luciferase, Activity Assay, Mutagenesis

Journal: International Journal of Biological Sciences

Article Title: Transcription Factor E2F4 Promote Proliferation, Migration, and Invasion of Gastric Cancer Cells by transcriptionally activating DSCC1

doi: 10.7150/ijbs.99590

Figure Lengend Snippet: Up-regulation of E2F4 in GC tissues and cells. (A) Comparison of F2F4 expression between tumor tissues and adjacent tissues from the TCGA database. (B) Comparison of E2F4 expression levels between high and low expression groups from the Kaplan-Meier Plotter. (C) Detection of E2F4 mRNA expression in 16 pairs of GC and corresponding adjacent tissues using RT-qPCR. (D) Detection of E2F4 protein expression in 16 pairs of GC and corresponding adjacent tissues using Western blot. (E) Analysis of E2F4 mRNA and protein expression in GES-1 and GC cell lines by RT-qPCR. (F) Analysis of E2F4 mRNA and protein expression in GES-1 and GC cell lines by Western blot. (G) Detection of E2F4 expression in 80 paired GC and adjacent non-tumor tissues by IHC. (H) Survival analysis of GC patients with varying levels of E2F4 expression. (*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001) - Significance levels denoted by asterisks.

Article Snippet: A portion of the lysates was designated as “Input.” The remaining lysates were incubated with E2F4 antibody (CST, #40291) and Protein G magnetic beads to form a DNA-antibody-magnetic bead complex.

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

Journal: International Journal of Biological Sciences

Article Title: Transcription Factor E2F4 Promote Proliferation, Migration, and Invasion of Gastric Cancer Cells by transcriptionally activating DSCC1

doi: 10.7150/ijbs.99590

Figure Lengend Snippet: Role of E2F4 in Promoting Malignancy in GC Cells. (A-B) Impact of E2F4 knockdown or overexpression on GC cell proliferation assessed through CCK-8 assay (A) and EdU assay (B) . (C) Influence of E2F4 knockdown or overexpression on GC cell cycle analyzed using flow cytometry. (D) Effect of E2F4 knockdown or overexpression on GC cell proliferation measured by colony formation assay. (E-F) Impact of E2F4 knockdown or overexpression on GC cell migration and invasion evaluated through transwell (E) and wound healing assay (F) . (G) GSEA analysis illustrating the correlation between E2F4 expression and the cell cycle pathway. (H) Alterations in cell cycle, proliferation, migration, and invasion-related proteins following E2F4 knockdown or overexpression analyzed by Western blot in GC cells. (*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001) - Significance levels denoted by asterisks.

Article Snippet: A portion of the lysates was designated as “Input.” The remaining lysates were incubated with E2F4 antibody (CST, #40291) and Protein G magnetic beads to form a DNA-antibody-magnetic bead complex.

Techniques: Knockdown, Over Expression, CCK-8 Assay, EdU Assay, Flow Cytometry, Colony Assay, Migration, Wound Healing Assay, Expressing, Western Blot

Journal: International Journal of Biological Sciences

Article Title: Transcription Factor E2F4 Promote Proliferation, Migration, and Invasion of Gastric Cancer Cells by transcriptionally activating DSCC1

doi: 10.7150/ijbs.99590

Figure Lengend Snippet: Role of E2F4 in Promoting Gastric Cancer Process In Vivo . (A) Inhibition of E2F4 suppressed the growth of subcutaneous xenograft tumors in nude mice. (B) Images of subcutaneous tumors. (C) Growth curve of subcutaneous tumors in nude mice. (D) Measurement of tumor weight post-sacrifice. (E) Western blot analysis of E2F4 expression in xenograft tumors. (*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001) - Significance levels denoted by asterisks.

Article Snippet: A portion of the lysates was designated as “Input.” The remaining lysates were incubated with E2F4 antibody (CST, #40291) and Protein G magnetic beads to form a DNA-antibody-magnetic bead complex.

Techniques: In Vivo, Inhibition, Western Blot, Expressing

Journal: International Journal of Biological Sciences

Article Title: Transcription Factor E2F4 Promote Proliferation, Migration, and Invasion of Gastric Cancer Cells by transcriptionally activating DSCC1

doi: 10.7150/ijbs.99590

Figure Lengend Snippet: DSCC1 is Crucial for E2F4-Mediated Malignant Phenotype in GC Cells. (A-B, D) Cell proliferation in NCI-N87 cells with E2F4 knockdown, control, and DSCC1 re-expression in E2F4-depleted cells, quantified using CCK-8 (A) , EdU (B) , and colony formation (D) assays. (C) Cell cycle analysis via flow cytometry in NCI-N87 cells with E2F4 knockdown, control, and DSCC1 re-expression in E2F4-depleted cells. (E) Assessment of cell migration and invasion through transwell assays in NCI-N87 cells with E2F4 knockdown, control, and DSCC1 re-expression in E2F4-depleted cells. (F) Western blot analysis of cell cycle, proliferation, migration, and invasion-related proteins in NCI-N87 cells with E2F4 knockdown, control, and DSCC1 re-expression in E2F4-depleted cells. (*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001) - Significance levels denoted by asterisks.

Article Snippet: A portion of the lysates was designated as “Input.” The remaining lysates were incubated with E2F4 antibody (CST, #40291) and Protein G magnetic beads to form a DNA-antibody-magnetic bead complex.

Techniques: Knockdown, Control, Expressing, CCK-8 Assay, Cell Cycle Assay, Flow Cytometry, Migration, Western Blot

Journal: iScience

Article Title: Sox9 mediates autophagy-dependent vascular smooth muscle cell phenotypic modulation and transplant arteriosclerosis

doi: 10.1016/j.isci.2022.105161

Figure Lengend Snippet: p27 associates with p130/E2F4 complex on Sox9 promoters (A) vSMCs were cotransfected with pGL3- Sox9 reporter construct and p27 overexpressing plasmid or empty vector for 48 h, Sox9 promoter activity was measured by dual luciferase assays (means ± SEM ∗p < 0.05. n = 3). (B) Dual luciferase assays were performed to evaluate Sox9 promoter activity in vSMCs cotransfected with pGL3- Sox9 reporter construct and si p27 for 48 h (means ± SEM ∗p < 0.05. n = 3). (C) Confirmation of the binding of p27 to Sox9 promoter in vSMCs by ChIP assay followed by qRT-PCR and agarose gel electrophoresis. An anti-IgG antibody was used as the negative control, an anti-Histone H3 antibody was used as the positive control and no antibody was added as the empty control (means ± SEM ∗∗p < 0.01, ∗∗∗p < 0.001. n = 4). (D) Coimmunoprecipitation (co-IP) of p27 and p130/E2f4 complex in vSMCs (n = 4). (E) ChIP of p130 and E2F4 protein in Sox9 promoter in vSMCs (means ± SEM ∗∗p < 0.01, ∗∗∗p < 0.001. n = 4). (F) vSMCs were cotransfected with pGL3- Sox9 reporter construct and p27 overexpressing plasmid or p130 siRNA or E2F4 siRNA, Sox9 promoter activity was measured by dual luciferase assays (means ± SEM ∗p < 0.05, ∗∗p < 0.01. n = 4). (G and H) qRT-PCR and western blotting analyses of Sox9 , Cnn1 , Acta2 and Tagln expression in vSMCs transfected with si p130 , si E2F4 or siNC for 48 h (means ± SEM ∗p < 0.05, ∗∗p < 0.01. n = 4).

Article Snippet: Rabbit anti-E2F4 antibody , Cell Signaling Technology , Cat#40291; RRID: AB_2799174.

Techniques: Construct, Plasmid Preparation, Activity Assay, Luciferase, Binding Assay, Quantitative RT-PCR, Agarose Gel Electrophoresis, Negative Control, Positive Control, Control, Co-Immunoprecipitation Assay, Western Blot, Expressing, Transfection

Journal: iScience

Article Title: Sox9 mediates autophagy-dependent vascular smooth muscle cell phenotypic modulation and transplant arteriosclerosis

doi: 10.1016/j.isci.2022.105161

Figure Lengend Snippet:

Article Snippet: Rabbit anti-E2F4 antibody , Cell Signaling Technology , Cat#40291; RRID: AB_2799174.

Techniques: Recombinant, CCK-8 Assay, Labeling, Sonication, Chromatin Immunoprecipitation, Enzyme-linked Immunosorbent Assay, Sequencing, Plasmid Preparation, Software, Imaging

Journal: Molecular Cell

Article Title: PAF remodels the DREAM complex to bypass cell quiescence and promote lung tumorigenesis

doi: 10.1016/j.molcel.2021.02.001

Figure Lengend Snippet:

Article Snippet: Rabbit anti-E2F4 , Bethyl , Cat# A302-134A; RRID: AB_1720353.

Techniques: Recombinant, shRNA, Software

Journal: Cancer Cell International

Article Title: Long non-coding RNA GAS5 accelerates oxidative stress in melanoma cells by rescuing EZH2-mediated CDKN1C downregulation

doi: 10.1186/s12935-020-01167-1

Figure Lengend Snippet: Overexpression of GAS5 prevents H3K27 trimethylation and upregulates CDKN1C expression to accelerate oxidative stress in melanoma by inhibiting EZH2. a Correlation analysis of lncRNA GAS5 expression and EZH2 expression. b RIP assay of the recruitment of transcription factor E2F4 by lncRNA GAS5. c ChIP assay of the enrichment of E2F4 in EZH2 promoter region. d RNA pull-down assay of the direct interaction of lncRNA GAS5 and E2F4. A375 cells were treated with oe-GAS5 alone or in the presence of oe-EZH2. e RT-qPCR assay of lncRNA GAS5 expression and mRNA expression of EZH2 and CDKN1C. f Western blot assay of the protein expression of E2F4, EZH2, H3K27me3 and CDKN1C. g Western blot assay of the protein expression of MDA5, IRE1α and SOD-1. h ELISA analysis of the content of ROS. i CCK-8 assay of cell viability. j Flow cytometric analysis of cell apoptosis. * p < 0.05, compared with the IgG group, cells transduced with oe-NC, Bio-probe NC group, cells transduced with oe-G-NC or cells transduced with oe-GAS5 + oe-E-NC. The above measurement data are expressed as mean ± standard deviation. The unpaired t -test is used for comparison of the data between two groups. Data among multiple groups are analyzed by one-way ANOVA, followed by Tukey’s post hoc test. The statistical analysis concerning time-based measurements within each group is realized using ANOVA of repeated measurements, followed by a Bonferroni’s post hoc test. Pearson correlation analysis is performed for correlation analysis

Article Snippet: The supernatants were collected and divided into three tubes, incubated overnight at 4 °C with the NC mouse antibody to IgG and target protein specific antibody, rabbit polyclonal antibody to EZH2 (ab195409, Abcam Inc., Cambridge, UK), rabbit monoclonal antibody to E2F4 (#40291, Cell Signaling Technologies, Beverly, MA, USA), rabbit monoclonal antibody to H3K27me3 (ab192985, Abcam Inc., Cambridge, UK).

Techniques: Over Expression, Expressing, Pull Down Assay, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Transduction, Standard Deviation, Comparison