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Genecopoeia human e2f2
Human E2f2, supplied by Genecopoeia, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+e2f2/Promoter+reporter+clone+for+Human+E2F2/pmc10104589-51-6-13
Average 94 stars, based on 1 article reviews

human e2f2 - by Bioz Stars, 2026-09

94/100 stars

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Plasmid Preparation:

Article Title: Engineered induced-pluripotent stem cell derived monocyte extracellular vesicles alter inflammation in HIV humanized mice
Article Snippet: Transfected EVs were purified with ExoQuick-TC (SBI) and stored at −80 °C until use [ ]. .. The plasmid containing 3’ UTR of human E2F2 (NM_004091.3, HMIT094972-MT05) was purchased from GeneCopoeia (Rockville, MD, USA). ..

Article Title: Engineered induced-pluripotent stem cell derived monocyte extracellular vesicles alter inflammation in HIV humanized mice.
Article Snippet: Transfected EVs were purified with ExoQuick-TC (SBI) and stored at -80 °C until use [Figure 1A]. .. Luciferase assay The plasmid containing 3’ UTR of human E2F2 (NM_004091.3, HMIT094972-MT05) was purchased from GeneCopoeia (Rockville, MD, USA). ..

Luciferase:

Image Search Results

The overexpression of C/EBPβ promoted the proliferation of HPCs. ( A ) The diagram shows that cells were disassociated by TrypLE at HPC D7 and then seeded at 4 × 10 4 cells/cm 2 on collagen type I coated plates. Simultaneously, PLV-C/EBPβ virus was administrated. Subsequently, cells were disassociated again and counted at HPC D8 and HPC D14, respectively. ( B ) Cell numbers were determined by cell counts for HPC on day 8 and for HPC on day 14. ( C ) GSEA results showed that cell cycle and E2F targets related genes were significantly enriched in HPCs transduced with PLV-C/EBPβ on day 14 when compared with those transduced with PLV-Vector on day 14. ( D ) Heat maps of selected proliferative genes and suppressor genes were differentially expressed between HPCs transduced with PLV-Vector or with PLV- C/EBPβ on day 14. ( E ) The KEGG pathways for the differentially expressed genes between HPCs transduced with PLV-Vector or with PLV- C/EBPβ on day 14. The principal pathways were the cell cycle and DNA replication. ( F ) Volcano plots showed significant changes in cell-cycle-related genes between HPCs transduced with PLV-Vector or with PLV- C/EBPβ on day 14. Volcano plot showed −log10 ( p -value) on the y-axis versus log2 (fold change) on the x-axis. Each point represented a different gene. ( G , H ) Relative expression of proliferative genes, including PCNA, CDC25C, CDC45L, MCM3, GINS1, and CCND1 ( F ) and E2F genes, including E2F1 and E2F2 ( H ) in HPCs transduced with PLV-Vector or with PLV- C/EBPβ on day 14. Data represent mean ± SEM with p -values indicated when significant. p < 0.05 *, p < 0.01 **, p < 0.001 ***, ns, not significant. Data are the three samples per group.

Journal: Cells

Article Title: C/EBPβ Coupled with E2F2 Promoted the Proliferation of hESC-Derived Hepatocytes through Direct Binding to the Promoter Regions of Cell-Cycle-Related Genes

doi: 10.3390/cells12030497

Figure Lengend Snippet: The overexpression of C/EBPβ promoted the proliferation of HPCs. ( A ) The diagram shows that cells were disassociated by TrypLE at HPC D7 and then seeded at 4 × 10 4 cells/cm 2 on collagen type I coated plates. Simultaneously, PLV-C/EBPβ virus was administrated. Subsequently, cells were disassociated again and counted at HPC D8 and HPC D14, respectively. ( B ) Cell numbers were determined by cell counts for HPC on day 8 and for HPC on day 14. ( C ) GSEA results showed that cell cycle and E2F targets related genes were significantly enriched in HPCs transduced with PLV-C/EBPβ on day 14 when compared with those transduced with PLV-Vector on day 14. ( D ) Heat maps of selected proliferative genes and suppressor genes were differentially expressed between HPCs transduced with PLV-Vector or with PLV- C/EBPβ on day 14. ( E ) The KEGG pathways for the differentially expressed genes between HPCs transduced with PLV-Vector or with PLV- C/EBPβ on day 14. The principal pathways were the cell cycle and DNA replication. ( F ) Volcano plots showed significant changes in cell-cycle-related genes between HPCs transduced with PLV-Vector or with PLV- C/EBPβ on day 14. Volcano plot showed −log10 ( p -value) on the y-axis versus log2 (fold change) on the x-axis. Each point represented a different gene. ( G , H ) Relative expression of proliferative genes, including PCNA, CDC25C, CDC45L, MCM3, GINS1, and CCND1 ( F ) and E2F genes, including E2F1 and E2F2 ( H ) in HPCs transduced with PLV-Vector or with PLV- C/EBPβ on day 14. Data represent mean ± SEM with p -values indicated when significant. p < 0.05 *, p < 0.01 **, p < 0.001 ***, ns, not significant. Data are the three samples per group.

Article Snippet: Subsequently, the aforementioned protein complexes were eluted from the beads, and then, agarose gel electrophoresis and Western blot were performed to transfer target proteins to PVDF membranes, followed by incubating with mouse anti-human C/EBPβ antibody (1:1000, santa cruz) or mouse anti-human E2F2 antibody (1:500, santa cruz).

Techniques: Over Expression, Transduction, Plasmid Preparation, Expressing

C/EBPβ Knockdown impaired hepatic differentiation and blocked proliferation of HPCs. ( A ) Representative images of cell morphologies of HPCs 2 days after si-RNA treatments on day 12 and cell numbers were counted in each group in ( A ). ( B ) Schematic illustration of si-RNA mediated C/EBPβ knockdown during the hepatic differentiation of hESCs. ( C ) Knockdown efficiency of C/EBPβ was detected by qRT-PCR on day 14 of HPCs. ( D ) ALB secretion was detected from the supernatants of cultured HPCs 24 h after siRNA (C/EBPβ) treatment. ( E ) Relative gene expression analysis of hepatocyte genes including AFP, ALB, ASGPR1, A1AT, HNF4A, and CEBPα in HPCs treated with si-NC or with si-C/EBPβ on day 14. ( F ) Heat map of selected genes, including hepatocyte genes, proliferative genes, and suppressor genes in HPCs treated with si-NC or with si-C/EBPβ on day 14. ( G ) The same number of LO2 cells were seeded, and then si-RNA mediated C/EBPβ knockdown was conducted. After 3 days of treatments, DAPI staining was performed to mark the nuclei, and then cell numbers were defined by ImageJ software. ( H ) Relative expression of proliferative genes including CDC25C, CDC45L, GINS1, MCM3, E2F1, and E2F2 in HPCs treated with si-NC or with si-C/EBPβ on day 14. ( I ) GSEA results showed that cell-cycle-related gene sets, such as MYC targets, cell-cycle literature, and mitosis targets bound by E2F, were significantly enriched in HPCs treated with si-NC when compared with those in HPCs treated with si-C/EBPβ. Data represent mean ± SEM with p -values indicated when significant. p < 0.05 *, p < 0.01 **, p < 0.0001 ****, ns, not significant. Data are the three samples per group.

Journal: Cells

Article Title: C/EBPβ Coupled with E2F2 Promoted the Proliferation of hESC-Derived Hepatocytes through Direct Binding to the Promoter Regions of Cell-Cycle-Related Genes

doi: 10.3390/cells12030497

Figure Lengend Snippet: C/EBPβ Knockdown impaired hepatic differentiation and blocked proliferation of HPCs. ( A ) Representative images of cell morphologies of HPCs 2 days after si-RNA treatments on day 12 and cell numbers were counted in each group in ( A ). ( B ) Schematic illustration of si-RNA mediated C/EBPβ knockdown during the hepatic differentiation of hESCs. ( C ) Knockdown efficiency of C/EBPβ was detected by qRT-PCR on day 14 of HPCs. ( D ) ALB secretion was detected from the supernatants of cultured HPCs 24 h after siRNA (C/EBPβ) treatment. ( E ) Relative gene expression analysis of hepatocyte genes including AFP, ALB, ASGPR1, A1AT, HNF4A, and CEBPα in HPCs treated with si-NC or with si-C/EBPβ on day 14. ( F ) Heat map of selected genes, including hepatocyte genes, proliferative genes, and suppressor genes in HPCs treated with si-NC or with si-C/EBPβ on day 14. ( G ) The same number of LO2 cells were seeded, and then si-RNA mediated C/EBPβ knockdown was conducted. After 3 days of treatments, DAPI staining was performed to mark the nuclei, and then cell numbers were defined by ImageJ software. ( H ) Relative expression of proliferative genes including CDC25C, CDC45L, GINS1, MCM3, E2F1, and E2F2 in HPCs treated with si-NC or with si-C/EBPβ on day 14. ( I ) GSEA results showed that cell-cycle-related gene sets, such as MYC targets, cell-cycle literature, and mitosis targets bound by E2F, were significantly enriched in HPCs treated with si-NC when compared with those in HPCs treated with si-C/EBPβ. Data represent mean ± SEM with p -values indicated when significant. p < 0.05 *, p < 0.01 **, p < 0.0001 ****, ns, not significant. Data are the three samples per group.

Techniques: Quantitative RT-PCR, Cell Culture, Expressing, Staining, Software

C/EBPβ coupled with E2F2 orchestrated cell proliferation of HPCs. ( A , B ) Western blot analysis of C/EBPβ on day 14 of HPCs after the knockdown or overexpression of C/EBPβ ( A ), and the statistical results ( B ). ( C , D ) Western blot analysis of representative protein expressions associated with cell proliferation after knockdown of C/EBPβ (C) , and corresponding statistical results ( D ). ( E , F ) Western blot analysis of representative protein expressions associated with cell proliferation after overexpression of C/EBPβ ( E ) and corresponding statistical results ( F ). ( G , H ) Co-immunoprecipitation (Co-IP) assays to validate interaction of C/EBPβ with E2F2 in HPCs on day 14 and LO2 cells. The cell lysates were subjected to immunoprecipitation with C/EBPβ antibody, and the resulting immunoprecipitants were analyzed in the immunoblot with E2F2 and C/EBPβ antibodies (IB: E2F2; IB: C/EBPβ). Input was also subjected to immunoblot to represent a positive control, and non-specific IgG represented as a negative control. Data represent mean ± SEM with p -values indicated when significant. p < 0.05 *, p < 0.01 **, ns, not significant. Data are the three samples per group.

Journal: Cells

Article Title: C/EBPβ Coupled with E2F2 Promoted the Proliferation of hESC-Derived Hepatocytes through Direct Binding to the Promoter Regions of Cell-Cycle-Related Genes

doi: 10.3390/cells12030497

Figure Lengend Snippet: C/EBPβ coupled with E2F2 orchestrated cell proliferation of HPCs. ( A , B ) Western blot analysis of C/EBPβ on day 14 of HPCs after the knockdown or overexpression of C/EBPβ ( A ), and the statistical results ( B ). ( C , D ) Western blot analysis of representative protein expressions associated with cell proliferation after knockdown of C/EBPβ (C) , and corresponding statistical results ( D ). ( E , F ) Western blot analysis of representative protein expressions associated with cell proliferation after overexpression of C/EBPβ ( E ) and corresponding statistical results ( F ). ( G , H ) Co-immunoprecipitation (Co-IP) assays to validate interaction of C/EBPβ with E2F2 in HPCs on day 14 and LO2 cells. The cell lysates were subjected to immunoprecipitation with C/EBPβ antibody, and the resulting immunoprecipitants were analyzed in the immunoblot with E2F2 and C/EBPβ antibodies (IB: E2F2; IB: C/EBPβ). Input was also subjected to immunoblot to represent a positive control, and non-specific IgG represented as a negative control. Data represent mean ± SEM with p -values indicated when significant. p < 0.05 *, p < 0.01 **, ns, not significant. Data are the three samples per group.

Techniques: Western Blot, Over Expression, Immunoprecipitation, Co-Immunoprecipitation Assay, Positive Control, Negative Control

C/EBPβ promoted the expression of proliferative genes by binding to promoter regions. ( A ) CUT&Tag data analysis for peak summit of relative genes at location map, the abscissa was the distance between peak summit and TSS site, and the ordinate was the number of peaks. ( B ) Pie chart of peak distribution on gene functional elements. ( C ) Top scoring and representative motif sequence of C/EBPβ binding sites. ( D ) KEGG enrichment bubble diagram to show the first 20 pathways with the smallest q values, pathway shown as the ordinate and rich factor shown as the abscissa (the number of differences in this pathway divided by all the numbers). The size indicated the number; the redder the color, the smaller the q value. ( E – I ) The peaks of individual proliferative genes, including CDC25C, CDC45L, PCNA, CDC7, and E2F2, were enriched in the transcriptional regulatory regions of their respective genes, and the peaks of some genes were enhanced after overexpression of C/EBPβ.

Journal: Cells

Article Title: C/EBPβ Coupled with E2F2 Promoted the Proliferation of hESC-Derived Hepatocytes through Direct Binding to the Promoter Regions of Cell-Cycle-Related Genes

doi: 10.3390/cells12030497

Figure Lengend Snippet: C/EBPβ promoted the expression of proliferative genes by binding to promoter regions. ( A ) CUT&Tag data analysis for peak summit of relative genes at location map, the abscissa was the distance between peak summit and TSS site, and the ordinate was the number of peaks. ( B ) Pie chart of peak distribution on gene functional elements. ( C ) Top scoring and representative motif sequence of C/EBPβ binding sites. ( D ) KEGG enrichment bubble diagram to show the first 20 pathways with the smallest q values, pathway shown as the ordinate and rich factor shown as the abscissa (the number of differences in this pathway divided by all the numbers). The size indicated the number; the redder the color, the smaller the q value. ( E – I ) The peaks of individual proliferative genes, including CDC25C, CDC45L, PCNA, CDC7, and E2F2, were enriched in the transcriptional regulatory regions of their respective genes, and the peaks of some genes were enhanced after overexpression of C/EBPβ.

Techniques: Expressing, Binding Assay, Functional Assay, Sequencing, Over Expression

Cell-cycle progression was promoted by C/EBPβ in LO2 cells. ( A ) The schematic diagram shows that C/EBPβ binds to CDC45L promoter regions to regulate luciferase expression. ( B ) Luciferase assays were carried out to verify the C/EBPβ binding activity at promoter regions of CDC45L gene in 293T cells and LO2 cells. ( C , D ) Lentivirus-mediated C/EBPβ knockdown with three pairs of sh-RNA (sh-a, sh-b, and sh-c) was performed to determine the expression of cell-cycle genes ( C ) and transcriptional factor E2Fs genes ( D ) by qRT-PCR assays. ( E , F ) Flow cytometry was performed to detect the proportion of cell-cycle phases after the treatment with sh-RNA (C/EBPβ) in LO2 cells ( E ), and the statistical results of proportion were depicted ( F ) by GraphPad Prism 8 software. ( G ) C/EBPβ coupled with E2F2 regulated cell-cycle gene expression in hepatic progenitor cells. C/EBPβ first bound to the regulatory region of cell-cycle-regulating transcription factor E2F2 to activate E2F2, then C/EBPβ coupled with E2F2 to promote the expression of cell-cycle genes (CDC45L, CDC25C, and PCNA) in hepatic progenitor cells. Data represent mean ± SEM with p -values indicated when significant. p < 0.05 *, p < 0.01 **, p < 0.001 ***, p < 0.0001 ****, ns, not significant. Data are the three samples per group.

Journal: Cells

Article Title: C/EBPβ Coupled with E2F2 Promoted the Proliferation of hESC-Derived Hepatocytes through Direct Binding to the Promoter Regions of Cell-Cycle-Related Genes

doi: 10.3390/cells12030497

Figure Lengend Snippet: Cell-cycle progression was promoted by C/EBPβ in LO2 cells. ( A ) The schematic diagram shows that C/EBPβ binds to CDC45L promoter regions to regulate luciferase expression. ( B ) Luciferase assays were carried out to verify the C/EBPβ binding activity at promoter regions of CDC45L gene in 293T cells and LO2 cells. ( C , D ) Lentivirus-mediated C/EBPβ knockdown with three pairs of sh-RNA (sh-a, sh-b, and sh-c) was performed to determine the expression of cell-cycle genes ( C ) and transcriptional factor E2Fs genes ( D ) by qRT-PCR assays. ( E , F ) Flow cytometry was performed to detect the proportion of cell-cycle phases after the treatment with sh-RNA (C/EBPβ) in LO2 cells ( E ), and the statistical results of proportion were depicted ( F ) by GraphPad Prism 8 software. ( G ) C/EBPβ coupled with E2F2 regulated cell-cycle gene expression in hepatic progenitor cells. C/EBPβ first bound to the regulatory region of cell-cycle-regulating transcription factor E2F2 to activate E2F2, then C/EBPβ coupled with E2F2 to promote the expression of cell-cycle genes (CDC45L, CDC25C, and PCNA) in hepatic progenitor cells. Data represent mean ± SEM with p -values indicated when significant. p < 0.05 *, p < 0.01 **, p < 0.001 ***, p < 0.0001 ****, ns, not significant. Data are the three samples per group.

Techniques: Luciferase, Expressing, Binding Assay, Activity Assay, Quantitative RT-PCR, Flow Cytometry, Software

Association of E2F2 expression and clinicopathological parameters in patients with CRC

Journal: Journal of Cancer

Article Title: Decreased E2F2 Expression Correlates with Poor Prognosis and Immune Infiltrates in Patients with Colorectal Cancer

doi: 10.7150/jca.61415

Figure Lengend Snippet: Association of E2F2 expression and clinicopathological parameters in patients with CRC

Article Snippet: After blocked with 5% skim milk in tris-buffered saline tween (TBST) at room temperature for 2 h, the membranes were probed with the primary antibodies: mouse anti-human E2F2 monoclonal antibody (55 kDa; 1:1000) (sc-9967, Santa Cruz), rabbit anti-human GAPDH antibody (36 kDa; 1:1000) (sc-47724, Santa Cruz) at 4 °C overnight.

Techniques: Expressing

Expression patterns of E2F2 mRNA in CRC from TIMER, TCGA and GEO database. (A) The E2F2 expression in different cancer types from the TIMER database. (B) E2F2 mRNA expression was significantly downregulated in CRC tumor tissues compared to normal tissues from the TCGA-COADREAD data sets. (C) E2F2 mRNA expression was significantly decreased in paired CRC tumor tissues compared to adjacent normal tissues from the TCGA-COADREAD data sets. (D) E2F2 mRNA expression was significantly lower in colon adenocarcinoma than normal tissues from the GSE20916 dataset. (E) E2F2 mRNA expression was significantly reduced in early stage CRC tumor tissues compared to normal tissues from the GSE9348 dataset. (F-I) The mRNA expression level of E2F2 was analyzed using the TCGA-COADREAD data sets according to (F) T, (G) N, (H) M and (I) pathological stage. ns, no significant difference; * , p < 0.05; **, p < 0.01; ***, p < 0.001.

Journal: Journal of Cancer

Article Title: Decreased E2F2 Expression Correlates with Poor Prognosis and Immune Infiltrates in Patients with Colorectal Cancer

doi: 10.7150/jca.61415

Figure Lengend Snippet: Expression patterns of E2F2 mRNA in CRC from TIMER, TCGA and GEO database. (A) The E2F2 expression in different cancer types from the TIMER database. (B) E2F2 mRNA expression was significantly downregulated in CRC tumor tissues compared to normal tissues from the TCGA-COADREAD data sets. (C) E2F2 mRNA expression was significantly decreased in paired CRC tumor tissues compared to adjacent normal tissues from the TCGA-COADREAD data sets. (D) E2F2 mRNA expression was significantly lower in colon adenocarcinoma than normal tissues from the GSE20916 dataset. (E) E2F2 mRNA expression was significantly reduced in early stage CRC tumor tissues compared to normal tissues from the GSE9348 dataset. (F-I) The mRNA expression level of E2F2 was analyzed using the TCGA-COADREAD data sets according to (F) T, (G) N, (H) M and (I) pathological stage. ns, no significant difference; * , p < 0.05; **, p < 0.01; ***, p < 0.001.

Techniques: Expressing

Logistic regression analysis of E2F2 expression associated with clinicopathological parameters in CRC

Journal: Journal of Cancer

Article Title: Decreased E2F2 Expression Correlates with Poor Prognosis and Immune Infiltrates in Patients with Colorectal Cancer

doi: 10.7150/jca.61415

Figure Lengend Snippet: Logistic regression analysis of E2F2 expression associated with clinicopathological parameters in CRC

Techniques: Expressing

E2F2 expression level was reduced in clinical CRC tissues. (A-D) Representative images of E2F2 protein expression level in paraffin-embedded (A) normal mucosa and CRC tissues with (B) pathological stage I, (C) pathological stage II and (D) pathological stage III. Scales represent 100 micron. Representative images with original magnificent at 200×. (E) Quantifications of the average optical density (AOD) for E2F2 protein expression in normal mucosa and CRC tissues with pathological stage I-III. (F) Western blotting analysis and (G) quantitative real-time polymerase chain reaction analysis of five paired samples of CRC tissues and adjacent normal tissues from randomly selected CRC patients. * , p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.

Journal: Journal of Cancer

Article Title: Decreased E2F2 Expression Correlates with Poor Prognosis and Immune Infiltrates in Patients with Colorectal Cancer

doi: 10.7150/jca.61415

Figure Lengend Snippet: E2F2 expression level was reduced in clinical CRC tissues. (A-D) Representative images of E2F2 protein expression level in paraffin-embedded (A) normal mucosa and CRC tissues with (B) pathological stage I, (C) pathological stage II and (D) pathological stage III. Scales represent 100 micron. Representative images with original magnificent at 200×. (E) Quantifications of the average optical density (AOD) for E2F2 protein expression in normal mucosa and CRC tissues with pathological stage I-III. (F) Western blotting analysis and (G) quantitative real-time polymerase chain reaction analysis of five paired samples of CRC tissues and adjacent normal tissues from randomly selected CRC patients. * , p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.

Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction

The diagnostic and prognostic value of E2F2 in CRC. (A-C) Kaplan-Meier survival curve analysis of OS, DFS and DSS showed that low E2F2 expression correlated to poor prognosis of CRC patients in a CRC cohort (GSE17537) from the PrognoScan database. (D-F) Survival curves showed OS, PFI and DSS rates of CRC patients with low or high E2F2 expression from the TCGA-COADREAD data sets. (G, H) ROC analysis illustrated that E2F2 expression accurately discriminated CRC tumor tissues from the normal tissues with an AUC of 0.865 (95% CI = 0.784-0.946) from GSE20916 data set and an AUC of 0.735 (95% CI = 0.672-0.798) from TCGA-COADREAD data sets. OS, overall survivial; DFS, disease free survival; DSS, disease specific survival; PFI, progress free interval; ROC: Receiver operating characteristic; AUC: area under the curve.

Journal: Journal of Cancer

Article Title: Decreased E2F2 Expression Correlates with Poor Prognosis and Immune Infiltrates in Patients with Colorectal Cancer

doi: 10.7150/jca.61415

Figure Lengend Snippet: The diagnostic and prognostic value of E2F2 in CRC. (A-C) Kaplan-Meier survival curve analysis of OS, DFS and DSS showed that low E2F2 expression correlated to poor prognosis of CRC patients in a CRC cohort (GSE17537) from the PrognoScan database. (D-F) Survival curves showed OS, PFI and DSS rates of CRC patients with low or high E2F2 expression from the TCGA-COADREAD data sets. (G, H) ROC analysis illustrated that E2F2 expression accurately discriminated CRC tumor tissues from the normal tissues with an AUC of 0.865 (95% CI = 0.784-0.946) from GSE20916 data set and an AUC of 0.735 (95% CI = 0.672-0.798) from TCGA-COADREAD data sets. OS, overall survivial; DFS, disease free survival; DSS, disease specific survival; PFI, progress free interval; ROC: Receiver operating characteristic; AUC: area under the curve.

Techniques: Diagnostic Assay, Expressing

Univariate and multivariate analysis of clinicopathological factors that correlate with OS of CRC patients

Journal: Journal of Cancer

Article Title: Decreased E2F2 Expression Correlates with Poor Prognosis and Immune Infiltrates in Patients with Colorectal Cancer

doi: 10.7150/jca.61415

Figure Lengend Snippet: Univariate and multivariate analysis of clinicopathological factors that correlate with OS of CRC patients

Techniques:

Go and KEGG enrichment analysis of genes related to E2F2 in CRC tissues in the TCGA-COADREAD data sets. (A-C) Go enrichment analysis showed the BP (biological processes), CC (cellular components), and MF (molecular function) of co-expressed genes with E2F2. (D) Significantly enriched KEGG terms obtained from KEGG enrichment analysis of co-expressed genes with E2F2.

Journal: Journal of Cancer

Article Title: Decreased E2F2 Expression Correlates with Poor Prognosis and Immune Infiltrates in Patients with Colorectal Cancer

doi: 10.7150/jca.61415

Figure Lengend Snippet: Go and KEGG enrichment analysis of genes related to E2F2 in CRC tissues in the TCGA-COADREAD data sets. (A-C) Go enrichment analysis showed the BP (biological processes), CC (cellular components), and MF (molecular function) of co-expressed genes with E2F2. (D) Significantly enriched KEGG terms obtained from KEGG enrichment analysis of co-expressed genes with E2F2.

Techniques:

Enrichment plots from the gene set enrichment analysis (GSEA). (A) ATR pathway, (B) ATM signalling pathway, (C) mismatch repair, (D) base excision repair, (E) homologous recomibination, (F) Fanconi Anemia pathway, (G) multicancer invasiveness signature, (H) stem cell up, and (I) mammary stem cell up were significantly enriched in E2F2-related CRC. NES, normalized enrichment scores; FDR, false discovery rate.

Journal: Journal of Cancer

Article Title: Decreased E2F2 Expression Correlates with Poor Prognosis and Immune Infiltrates in Patients with Colorectal Cancer

doi: 10.7150/jca.61415

Figure Lengend Snippet: Enrichment plots from the gene set enrichment analysis (GSEA). (A) ATR pathway, (B) ATM signalling pathway, (C) mismatch repair, (D) base excision repair, (E) homologous recomibination, (F) Fanconi Anemia pathway, (G) multicancer invasiveness signature, (H) stem cell up, and (I) mammary stem cell up were significantly enriched in E2F2-related CRC. NES, normalized enrichment scores; FDR, false discovery rate.

Techniques:

Association analysis of E2F2 gene expression and immune infiltration. (A) The association between E2F2 expression and 24 tumor-infiltrating lymphocytes. (B-G) The correlation of E2F2 expression with immune infiltration level of (B) Th2 cells, (C) aDC cells, (D) Th17 cells, (E) NK CD56dim cells, (F) T helper cells, (G) pDC cells.

Journal: Journal of Cancer

Article Title: Decreased E2F2 Expression Correlates with Poor Prognosis and Immune Infiltrates in Patients with Colorectal Cancer

doi: 10.7150/jca.61415

Figure Lengend Snippet: Association analysis of E2F2 gene expression and immune infiltration. (A) The association between E2F2 expression and 24 tumor-infiltrating lymphocytes. (B-G) The correlation of E2F2 expression with immune infiltration level of (B) Th2 cells, (C) aDC cells, (D) Th17 cells, (E) NK CD56dim cells, (F) T helper cells, (G) pDC cells.

Techniques: Gene Expression, Expressing

Expression of E2F2 in synovial tissues (STs). A , E2F2 mRNA expression in ST from patients with rheumatoid arthritis (RA; n = 10) and osteoarthritis (OA; n = 10) was determined by real-time PCR and calculated as 2−∆∆Ct, with the GAPDH gene used as an endogenous control. B , Expression of the E2F2 protein in ST from RA(n = 10) and OA(n = 10) was evaluated by western blot analysis, with the β-actin protein used as a loading control. Values are expressed as mean ± SD of the mean and at least 3 independent experiments were performed; *p < 0.05.

Journal: Scientific Reports

Article Title: A critical role of E2F transcription factor 2 in proinflammatory cytokines-dependent proliferation and invasiveness of fibroblast-like synoviocytes in rheumatoid Arthritis

doi: 10.1038/s41598-018-20782-7

Figure Lengend Snippet: Expression of E2F2 in synovial tissues (STs). A , E2F2 mRNA expression in ST from patients with rheumatoid arthritis (RA; n = 10) and osteoarthritis (OA; n = 10) was determined by real-time PCR and calculated as 2−∆∆Ct, with the GAPDH gene used as an endogenous control. B , Expression of the E2F2 protein in ST from RA(n = 10) and OA(n = 10) was evaluated by western blot analysis, with the β-actin protein used as a loading control. Values are expressed as mean ± SD of the mean and at least 3 independent experiments were performed; *p < 0.05.

Article Snippet: Anti-human E2F2 antibody was diluted at 1:1000 (Millipore, USA), and anti-human β-actin antibody diluted at 1:1000 (Beoytime, China).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control, Western Blot

Effect of E2F2 on RASFs proliferation, invasion, migration. A-B , RASFs were transiently transfected with siRNA targeting E2F2.Resultant expression was detected using qPCR ( A ) and western blot ( B ). The E2F2 mRNA level and protein levels were both normalized to β-actin. ( C) , MTS assay was used to assess cell proliferation. ( D ), The invasive ability was performed by transwell assay and the average number of cells that invaded through the filter was quantified. E-F , Migration capacity of RASFs 24 h after silencing E2F2 by transwell( E ) and scratching( F ). Results of statistical analyses of three independent determinations. NC: negative control; MOCK: transfection-reagent control. Values are expressed as mean ± SD of the mean and at least 3 independent experiments were performed; *p < 0.05.

Journal: Scientific Reports

Article Title: A critical role of E2F transcription factor 2 in proinflammatory cytokines-dependent proliferation and invasiveness of fibroblast-like synoviocytes in rheumatoid Arthritis

doi: 10.1038/s41598-018-20782-7

Figure Lengend Snippet: Effect of E2F2 on RASFs proliferation, invasion, migration. A-B , RASFs were transiently transfected with siRNA targeting E2F2.Resultant expression was detected using qPCR ( A ) and western blot ( B ). The E2F2 mRNA level and protein levels were both normalized to β-actin. ( C) , MTS assay was used to assess cell proliferation. ( D ), The invasive ability was performed by transwell assay and the average number of cells that invaded through the filter was quantified. E-F , Migration capacity of RASFs 24 h after silencing E2F2 by transwell( E ) and scratching( F ). Results of statistical analyses of three independent determinations. NC: negative control; MOCK: transfection-reagent control. Values are expressed as mean ± SD of the mean and at least 3 independent experiments were performed; *p < 0.05.

Article Snippet: Anti-human E2F2 antibody was diluted at 1:1000 (Millipore, USA), and anti-human β-actin antibody diluted at 1:1000 (Beoytime, China).

Techniques: Migration, Transfection, Expressing, Western Blot, MTS Assay, Transwell Assay, Negative Control, Control

Effects of cytokines on expression of E2F2 and signaling pathways involved in the stimulation. A–C , Effect of signaling pathway inhibitors on cytokines induction of E2F2. RASFs were treated for 2 hours with IL-6( A ), TNF-α ( B ), and LPS( C ) separately, or in combination with pyrrolidine dithiocarbamate (PDTC), PD98059, or Stattic respectively for 24 h. mRNA and protein expression of E2F2 in cell culture were evaluated by RT-qPCR and western blot respectively. ( D ), Schematic representation of E2F2 promoters, primers for the ChIP assay, NF-кB binding motif in E2F2 promoter. ( E ), Binding of the E2F2 gene promoter in RASFs in vitro . ChIP-PCR was used to reveal whether p65 of NF-κB could bind to the promoter of the E2F2 gene in RASFs in vitro , and the effect of TNF-α stimulation on the binding was assayed using immunoprecipitated chromatin with anti-p65 antibody; (Input, chromatin input before immunoprecipitation; IgG, immunoprecipitated chromatin with control IgG;Anti-p50, immunoprecipitated chromatin with anti-p50 antibody; Anti-p65, immunoprecipitated chromatin with anti-p65 antibody). ( F ), HEK293T cells stimulated with TNF-α, the luciferase reporter activity of NF-κB was then monitored. The data shown are means ± s.e.m and representation of six independent experiments. *P < 0.05, **P < 0.01, Values are expressed as mean ± SD of the mean and at least 3 independent experiments were performed.

Journal: Scientific Reports

Article Title: A critical role of E2F transcription factor 2 in proinflammatory cytokines-dependent proliferation and invasiveness of fibroblast-like synoviocytes in rheumatoid Arthritis

doi: 10.1038/s41598-018-20782-7

Figure Lengend Snippet: Effects of cytokines on expression of E2F2 and signaling pathways involved in the stimulation. A–C , Effect of signaling pathway inhibitors on cytokines induction of E2F2. RASFs were treated for 2 hours with IL-6( A ), TNF-α ( B ), and LPS( C ) separately, or in combination with pyrrolidine dithiocarbamate (PDTC), PD98059, or Stattic respectively for 24 h. mRNA and protein expression of E2F2 in cell culture were evaluated by RT-qPCR and western blot respectively. ( D ), Schematic representation of E2F2 promoters, primers for the ChIP assay, NF-кB binding motif in E2F2 promoter. ( E ), Binding of the E2F2 gene promoter in RASFs in vitro . ChIP-PCR was used to reveal whether p65 of NF-κB could bind to the promoter of the E2F2 gene in RASFs in vitro , and the effect of TNF-α stimulation on the binding was assayed using immunoprecipitated chromatin with anti-p65 antibody; (Input, chromatin input before immunoprecipitation; IgG, immunoprecipitated chromatin with control IgG;Anti-p50, immunoprecipitated chromatin with anti-p50 antibody; Anti-p65, immunoprecipitated chromatin with anti-p65 antibody). ( F ), HEK293T cells stimulated with TNF-α, the luciferase reporter activity of NF-κB was then monitored. The data shown are means ± s.e.m and representation of six independent experiments. *P < 0.05, **P < 0.01, Values are expressed as mean ± SD of the mean and at least 3 independent experiments were performed.

Article Snippet: Anti-human E2F2 antibody was diluted at 1:1000 (Millipore, USA), and anti-human β-actin antibody diluted at 1:1000 (Beoytime, China).

Techniques: Expressing, Protein-Protein interactions, Cell Culture, Quantitative RT-PCR, Western Blot, Binding Assay, In Vitro, Immunoprecipitation, Control, Luciferase, Activity Assay

$Effect of cytokines on E2F2 nuclear translocation in RASFs and subsequent effects of nuclear translocation. ( A ), Nuclear and cytoplasmic proteins were fractionally extracted from RASFs stimulated with IL-6, TNF-α, and LPS for 6 h. Effects of cytokines on nuclear translocation of E2F2 were determined by Western blot. (Lamin A/C: reference for nuclear extraction; β-actin: reference for cytoplasmic extraction; control: C: Cytoplasmic extraction; N: Nuclear Extraction). ( B ), Nuclear translocation of E2F2 observed using confocal fluorescences microscopy. E2F2 was detected using anti-E2F2 antibody, and Alexa Fluor 488-conjugated IgG. Nuclei were stained with 4′,6′-diamidino-2-phenylinndole (DAPI, blue). The images showed more E2F2 translocation into the nucleus after stimulation with LPS (I), TNF-α(II), and IL-6 (III). ( C ), Measurement of cytokine levels affected by E2F2. mRNA expression and secretion of IL-6, IL-1α, IL-1β, and TNF-α was determined by real-time PCR and ELISA respectively. ( D ), Schematic representation of IL-6 promoters, primers for the ChIP assay, E2F2 binding motif in IL-6 promoter. ( E ), ChIP-PCR was used to determine whether E2F2 affect IL-6 secretion by binding to its promoter directly. Effects of TNF-α and LPS on the bindation between E2F2 and the promoter of IL-6 were also investigated. (Input, chromatin input before immunoprecipitation; Anti-E2F2: immunoprecipitated chromatin with anti-E2F2 antibody; IgG, immunoprecipitated chromatin with control IgG.) (F) , HEK293T cells stimulated with LPS and TNF-α, the luciferase reporter activity of E2F2 was then monitored. Values are expressed as mean ± SD of the mean and at least 3 independent experiments were performed. *P < 0.05, **P < 0.01.$

Journal: Scientific Reports

Article Title: A critical role of E2F transcription factor 2 in proinflammatory cytokines-dependent proliferation and invasiveness of fibroblast-like synoviocytes in rheumatoid Arthritis

doi: 10.1038/s41598-018-20782-7

Figure Lengend Snippet: Effect of cytokines on E2F2 nuclear translocation in RASFs and subsequent effects of nuclear translocation. ( A ), Nuclear and cytoplasmic proteins were fractionally extracted from RASFs stimulated with IL-6, TNF-α, and LPS for 6 h. Effects of cytokines on nuclear translocation of E2F2 were determined by Western blot. (Lamin A/C: reference for nuclear extraction; β-actin: reference for cytoplasmic extraction; control: C: Cytoplasmic extraction; N: Nuclear Extraction). ( B ), Nuclear translocation of E2F2 observed using confocal fluorescences microscopy. E2F2 was detected using anti-E2F2 antibody, and Alexa Fluor 488-conjugated IgG. Nuclei were stained with 4′,6′-diamidino-2-phenylinndole (DAPI, blue). The images showed more E2F2 translocation into the nucleus after stimulation with LPS (I), TNF-α(II), and IL-6 (III). ( C ), Measurement of cytokine levels affected by E2F2. mRNA expression and secretion of IL-6, IL-1α, IL-1β, and TNF-α was determined by real-time PCR and ELISA respectively. ( D ), Schematic representation of IL-6 promoters, primers for the ChIP assay, E2F2 binding motif in IL-6 promoter. ( E ), ChIP-PCR was used to determine whether E2F2 affect IL-6 secretion by binding to its promoter directly. Effects of TNF-α and LPS on the bindation between E2F2 and the promoter of IL-6 were also investigated. (Input, chromatin input before immunoprecipitation; Anti-E2F2: immunoprecipitated chromatin with anti-E2F2 antibody; IgG, immunoprecipitated chromatin with control IgG.) (F) , HEK293T cells stimulated with LPS and TNF-α, the luciferase reporter activity of E2F2 was then monitored. Values are expressed as mean ± SD of the mean and at least 3 independent experiments were performed. *P < 0.05, **P < 0.01.

Article Snippet: Anti-human E2F2 antibody was diluted at 1:1000 (Millipore, USA), and anti-human β-actin antibody diluted at 1:1000 (Beoytime, China).

Techniques: Translocation Assay, Western Blot, Extraction, Control, Microscopy, Staining, Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Binding Assay, Immunoprecipitation, Luciferase, Activity Assay

Effect of E2F2 on IL-6 expression and correlations between serum E2F2 and cytokine levels in patients with rheumatoid arthritis. ( A ), Effect of siE2F2 on TNF-α stimulated IL-6 expression detected by RT-qPCR (left) and Western blot (right). ( B ), Correlations between mRNA levels of E2F2 and IL-6 or TNF-α secreted into the supernatant of RASFs. qRT-PCR was used to quantify mRNA levels of E 2 F 2 , and ELISA was performed to quantify the levels and IL-6 or TNF-α respectively secreted into the supernatant of RASFs(n = 15). Pearson correlation analysis was used to assay the correlations. Values are expressed as mean ± SD of the mean and at least 3 independent experiments were performed. *P < 0.05. **P < 0.01.

Journal: Scientific Reports

Article Title: A critical role of E2F transcription factor 2 in proinflammatory cytokines-dependent proliferation and invasiveness of fibroblast-like synoviocytes in rheumatoid Arthritis

doi: 10.1038/s41598-018-20782-7

Figure Lengend Snippet: Effect of E2F2 on IL-6 expression and correlations between serum E2F2 and cytokine levels in patients with rheumatoid arthritis. ( A ), Effect of siE2F2 on TNF-α stimulated IL-6 expression detected by RT-qPCR (left) and Western blot (right). ( B ), Correlations between mRNA levels of E2F2 and IL-6 or TNF-α secreted into the supernatant of RASFs. qRT-PCR was used to quantify mRNA levels of E 2 F 2 , and ELISA was performed to quantify the levels and IL-6 or TNF-α respectively secreted into the supernatant of RASFs(n = 15). Pearson correlation analysis was used to assay the correlations. Values are expressed as mean ± SD of the mean and at least 3 independent experiments were performed. *P < 0.05. **P < 0.01.

Article Snippet: Anti-human E2F2 antibody was diluted at 1:1000 (Millipore, USA), and anti-human β-actin antibody diluted at 1:1000 (Beoytime, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay

E2F2 is a direct target of miR-218. (A) Predicted binding sites for the seed sequences of miR-218 and miR-520a in the E2F2 3′-UTR and the sites of target mutagenesis are presented. (B) A dual luciferase assay was conducted in order to confirm the direct regulation of miR-218 on the E2F2 3′-UTR. miR-218 and miR-520a regulate E2F2 expression at the (C) mRNA and (D) protein levels in Huh7 and MHCC-97H cells. Values are presented as the mean ± standard deviation (n=3). * P<0.05, ** P<0.01, vs. NC. miR-218, microRNA-218; UTR, untranslated region; NC, negative control; Wt, wild-type; mt, mutant.

Journal: Molecular Medicine Reports

Article Title: MicroRNA-218 and microRNA-520a inhibit cell proliferation by downregulating E2F2 in hepatocellular carcinoma

doi: 10.3892/mmr.2015.3516

Figure Lengend Snippet: E2F2 is a direct target of miR-218. (A) Predicted binding sites for the seed sequences of miR-218 and miR-520a in the E2F2 3′-UTR and the sites of target mutagenesis are presented. (B) A dual luciferase assay was conducted in order to confirm the direct regulation of miR-218 on the E2F2 3′-UTR. miR-218 and miR-520a regulate E2F2 expression at the (C) mRNA and (D) protein levels in Huh7 and MHCC-97H cells. Values are presented as the mean ± standard deviation (n=3). * P<0.05, ** P<0.01, vs. NC. miR-218, microRNA-218; UTR, untranslated region; NC, negative control; Wt, wild-type; mt, mutant.

Article Snippet: Following blocking with 5% skimmed milk, the membranes were incubated with rabbit anti-human E2F2 polyclonal antibody (1:200; sc-632; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) overnight at 4°C.

Techniques: Binding Assay, Mutagenesis, Luciferase, Expressing, Standard Deviation, Negative Control

BCL2L11(BIM)-P1 activity is regulated by E2F . For simplicity the alias BIM is used in the figure instead of the approved gene symbol BIM. A . Inducibility of endogenous E1 and E2-bearing BIM transcripts by transiently transfected E2F expression construct in HEK293 cells; real-time experiments were performed as previously in cells transfected with either pBst or an E2F expression construct, and the fold inducibility (relatively to pBst transfected cells) of individual transcripts by the E2F expression plasmid is presented; the results confirm reporter data and show that the expression of endogenous BIM transcripts derived from P1 can be induced by E2F. *p < 0.01, Anova: single factor. B . Analysis of the effects of E2F overerexpression on pBIM-P1-1918(+) activity in HEK293 cells. A concentration-dependent inducibility of P1 activity is observed; by contrast, a version of P1 comprising a 5' deletion of 1300 bp results in a promoter construct with a basal activity comparable to P1-1918, but without the capacity for E2F-mediated activation, suggesting the presence of E2F responsive elements in the deleted region. *p < 0.01, Anova: single factor.

Journal: BMC Molecular Biology

Article Title: Identification of a candidate alternative promoter region of the human Bcl2L11 (Bim) gene

doi: 10.1186/1471-2199-9-56

Figure Lengend Snippet: BCL2L11(BIM)-P1 activity is regulated by E2F . For simplicity the alias BIM is used in the figure instead of the approved gene symbol BIM. A . Inducibility of endogenous E1 and E2-bearing BIM transcripts by transiently transfected E2F expression construct in HEK293 cells; real-time experiments were performed as previously in cells transfected with either pBst or an E2F expression construct, and the fold inducibility (relatively to pBst transfected cells) of individual transcripts by the E2F expression plasmid is presented; the results confirm reporter data and show that the expression of endogenous BIM transcripts derived from P1 can be induced by E2F. *p < 0.01, Anova: single factor. B . Analysis of the effects of E2F overerexpression on pBIM-P1-1918(+) activity in HEK293 cells. A concentration-dependent inducibility of P1 activity is observed; by contrast, a version of P1 comprising a 5' deletion of 1300 bp results in a promoter construct with a basal activity comparable to P1-1918, but without the capacity for E2F-mediated activation, suggesting the presence of E2F responsive elements in the deleted region. *p < 0.01, Anova: single factor.

Article Snippet: HEK293 cells were cotransfected with 0.05 μg of reporter plasmid, 0.01 μg of Renilla vector (pRL-TK, Promega), 0.01 μg of empty plasmid Bluescript (Stratagene, Inc.) or 0.1 or 0.05 μg Human E2F expression construct (Origene, Inc.) in 95 μl of serum-free medium per well in 96-well plate using Lipofectamine 2000 [ ].

Techniques: Activity Assay, Transfection, Expressing, Construct, Plasmid Preparation, Derivative Assay, Concentration Assay, Activation Assay

Effects of E2F on BCL2L11-P1 constructs . For simplicity the alias BIM is used in the figure instead of the approved gene symbol BCL2L11. A . Sequence of the -1918 P1 region showing the position of 5' deletion variants. Forward primers used to generate the various P1 5' deletion variants are indicated (arrows). The putative E2F binding site is boxed, and the transcript initiation site is indicated (+1). B . Analysis of the effects of E2F over-expression on pBIM-P1 and 5'deletion variants activity in HEK293 cells. A concentration-dependent inducibility of P1 activity is observed in construct pBIM-P1-1918, -1565 and -1326, which contain a putative E2F responsive element. By contrast, pBIM-P1-1284 and -940 display basal activities comparable to pBCL2L11-P1-1918, but without the capacity for E2F-mediated response, suggesting a loss of E2F responsive element(s). *p < 0.05 and **p < 0.01, Anova: single factor.

Journal: BMC Molecular Biology

Article Title: Identification of a candidate alternative promoter region of the human Bcl2L11 (Bim) gene

doi: 10.1186/1471-2199-9-56

Figure Lengend Snippet: Effects of E2F on BCL2L11-P1 constructs . For simplicity the alias BIM is used in the figure instead of the approved gene symbol BCL2L11. A . Sequence of the -1918 P1 region showing the position of 5' deletion variants. Forward primers used to generate the various P1 5' deletion variants are indicated (arrows). The putative E2F binding site is boxed, and the transcript initiation site is indicated (+1). B . Analysis of the effects of E2F over-expression on pBIM-P1 and 5'deletion variants activity in HEK293 cells. A concentration-dependent inducibility of P1 activity is observed in construct pBIM-P1-1918, -1565 and -1326, which contain a putative E2F responsive element. By contrast, pBIM-P1-1284 and -940 display basal activities comparable to pBCL2L11-P1-1918, but without the capacity for E2F-mediated response, suggesting a loss of E2F responsive element(s). *p < 0.05 and **p < 0.01, Anova: single factor.

Techniques: Construct, Sequencing, Binding Assay, Over Expression, Activity Assay, Concentration Assay

E2F binding to a candidate E2F binding site in BCL2L11-P1 . EMSA on the candidate E2F binding site (CTCTGCGCGCCAGAGG). HEK293 cells were transfected with the E2F expression construct, and nuclear extracts were assayed for capacity for specific binding to the candidate E2F binding site through competition with a specific (E2F) or unspecific (NF-κB) unlabelled competitor double stranded (ds) oligonucleotide. A specific shift is identified (arrow) upon transfection with the E2F expression plasmid, which can only be competed with a specific ds oligonucleotide.

Journal: BMC Molecular Biology

Article Title: Identification of a candidate alternative promoter region of the human Bcl2L11 (Bim) gene

doi: 10.1186/1471-2199-9-56

Figure Lengend Snippet: E2F binding to a candidate E2F binding site in BCL2L11-P1 . EMSA on the candidate E2F binding site (CTCTGCGCGCCAGAGG). HEK293 cells were transfected with the E2F expression construct, and nuclear extracts were assayed for capacity for specific binding to the candidate E2F binding site through competition with a specific (E2F) or unspecific (NF-κB) unlabelled competitor double stranded (ds) oligonucleotide. A specific shift is identified (arrow) upon transfection with the E2F expression plasmid, which can only be competed with a specific ds oligonucleotide.

Techniques: Binding Assay, Transfection, Expressing, Construct, Plasmid Preparation

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