Journal: Journal of Cellular and Molecular Medicine

Article Title: Classification of Paediatric Celiac Disease Using RNA Sequencing and Real‐Time PCR of Duodenal Biomarkers

doi: 10.1111/jcmm.70854

Figure Lengend Snippet: (a–d) Expression of (a) CYP3A4 , (b) LCT , (c) CXCL11 , and (d) E2F5 , normalised to the lowest expression in the whole dataset and presented on a log 2 scale. The grey marker in group BiopsyDiag represents the misclassified biopsy from case 51, and the grey marker in group TGdiag represents the misclassified biopsy from case 55. From left to right in each plot, the groups are: Not CD, Normalised CD, Potential CD without progression, Potential CD with progression, Potential CD with high anti‐TG2 and progression, Biopsy‐based CD diagnosis, Anti‐TG2 based CD diagnosis, and Marsh 3 histopathology and negative anti‐TG2.

Article Snippet: E2F5 , E2F transcription factor 5 , 8q21.2 , Hs00231092_m1.

Techniques: Expressing, Marker, Biomarker Discovery, Histopathology

Journal: Blood and Lymphatic Cancer: Targets and Therapy

Article Title: Crosstalk Between FOXN3 and E2F5 Reveals a Novel Tumor Suppressive Pathway in Acute Myeloid Leukemia via MAPK Signaling: Implications for Potential Future Targeted Therapy

doi: 10.2147/BLCTT.S571966

Figure Lengend Snippet: Identification of E2F5 as a transcriptional target of FOXN3. (A ) Genomic distribution of FOXN3 binding sites, represented through pie charts. ( B ) Distribution of binding sites relative to transcription initiation peaks within 5 kb upstream and downstream of the transcription start site (TSS). ( C and D ) RT-qPCR ( C ) and Western blotting ( D ) analyses of E2F5 expression in THP1 cells following FOXN3 overexpression. ( E ) Scatter plot illustrating the correlation between FOXN3 and E2F5 expression levels in AML patient samples from the GSE13159 dataset, indicating a negative correlation between the two genes. ( F ) Scatter plot confirming the negative correlation between FOXN3 and E2F5 expression in AML patient samples collected from our institution. ( G ) Visualization of representative FOXN3 binding peaks in the genome browser using karyoploteR, specifically in the E2F5 promoter region. The tracks display both input and immunoprecipitation (IP) data for the Kasumi1 and THP1 cell lines. The highlighted region denotes the E2F5 promoter area, where significant binding peaks were detected in the ChIP-seq experiment. ( H ) ChIP-PCR analysis demonstrating FOXN3 binding to the E2F5 promoter. Significant enrichment of FOXN3 binding is observed at the E2F5 promoter compared to the IgG control, with the SIRT6 promoter serving as a positive control ( P = 0.001). ( I ) Results from the luciferase reporter assay illustrating the repressive effect of FOXN3 on E2F5 promoter activity. The relative luciferase activity is significantly lower in the presence of FOXN3 ( P < 0.01). Data are presented as mean ± SD of three independent experiments. Statistical significance is indicated as follows: * P < 0.05, ** P < 0.01.

Article Snippet: The primary antibodies used were as follows: FOXN3 (ab129453, 1:2500, Abcam, UK), E2F5 (A16042, 1:500, ABclonal, China), CDC42 (2466T, 1:1000, Cell Signaling Technology, MA), HDAC1 (34589T, 1:1000, Cell Signaling Technology, MA), PCNA (WL03213, 1:500, Wanleibio, China), CyclinE1 (WL01072, 1:1000, Wanleibio, China), CDK2 (WL01543, 1:500, Wanleibio, China), CyclinD1 (WL01435a, 1:400, Wanleibio, China), CDK6 (WL03460, 1:500, Wanleibio, China), Bcl-2 (WL01556, 1:1000, Wanleibio, China), EZH2 (WL00795, 1:1000, Wanleibio, China), p-ERK (WLP1512, 1:1000, Wanleibio, China), ERK (WL01864, 1:1000, Wanleibio, China), p-38 (WL00764, 1:1000, Wanleibio, China), and GAPDH (5174, 1:1000, Cell Signaling Technology, MA).

Techniques: Binding Assay, Quantitative RT-PCR, Western Blot, Expressing, Over Expression, Immunoprecipitation, ChIP-sequencing, Control, Positive Control, Luciferase, Reporter Assay, Activity Assay

Journal: Blood and Lymphatic Cancer: Targets and Therapy

Article Title: Crosstalk Between FOXN3 and E2F5 Reveals a Novel Tumor Suppressive Pathway in Acute Myeloid Leukemia via MAPK Signaling: Implications for Potential Future Targeted Therapy

doi: 10.2147/BLCTT.S571966

Figure Lengend Snippet: Aberrant upregulation of E2F5 expression and its association with poor outcomes in AML. ( A ) RT-qPCR analysis showing elevated E2F5 mRNA expression in AML patients compared to healthy controls. ( B ) E2F5 mRNA levels are significantly higher in AML compared to hematopoietic stem cells (HSCs), as demonstrated using the online Bloodpool dataset. ( C ) RT-qPCR analysis revealing increased E2F5 mRNA expression in AML cell lines compared to control cell lines. ( D ) Correlation between higher E2F5 expression and shorter overall survival (OS) in AML, determined using the median value as the cutoff to distinguish between high and low E2F5 expression levels from the online GEPIA dataset. Data are presented as mean ± SD of three independent experiments. Statistical significance is indicated as follows:* P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: The primary antibodies used were as follows: FOXN3 (ab129453, 1:2500, Abcam, UK), E2F5 (A16042, 1:500, ABclonal, China), CDC42 (2466T, 1:1000, Cell Signaling Technology, MA), HDAC1 (34589T, 1:1000, Cell Signaling Technology, MA), PCNA (WL03213, 1:500, Wanleibio, China), CyclinE1 (WL01072, 1:1000, Wanleibio, China), CDK2 (WL01543, 1:500, Wanleibio, China), CyclinD1 (WL01435a, 1:400, Wanleibio, China), CDK6 (WL03460, 1:500, Wanleibio, China), Bcl-2 (WL01556, 1:1000, Wanleibio, China), EZH2 (WL00795, 1:1000, Wanleibio, China), p-ERK (WLP1512, 1:1000, Wanleibio, China), ERK (WL01864, 1:1000, Wanleibio, China), p-38 (WL00764, 1:1000, Wanleibio, China), and GAPDH (5174, 1:1000, Cell Signaling Technology, MA).

Techniques: Expressing, Quantitative RT-PCR, Control

Journal: Blood and Lymphatic Cancer: Targets and Therapy

Article Title: Crosstalk Between FOXN3 and E2F5 Reveals a Novel Tumor Suppressive Pathway in Acute Myeloid Leukemia via MAPK Signaling: Implications for Potential Future Targeted Therapy

doi: 10.2147/BLCTT.S571966

Figure Lengend Snippet: Inhibition of cell proliferation, induction of apoptosis, and cell cycle arrest following E2F5 knockdown in AML cells. ( A ) CCK-8 assay was conducted to assess the proliferation of Kasumi1 and THP1 cells following E2F5 knockdown. ( B ) Flow cytometry analysis evaluated the induction of apoptosis in Kasumi1 and THP1 cells after E2F5 knockdown. ( C ) Cell cycle analysis was performed on Kasumi1 and THP1 cells with E2F5 knockdown using flow cytometry. Data are presented as mean ± SD of three independent experiments. Statistical significance is indicated as follows: * P < 0.05, ** P < 0.01, and **** P < 0.0001.

Article Snippet: The primary antibodies used were as follows: FOXN3 (ab129453, 1:2500, Abcam, UK), E2F5 (A16042, 1:500, ABclonal, China), CDC42 (2466T, 1:1000, Cell Signaling Technology, MA), HDAC1 (34589T, 1:1000, Cell Signaling Technology, MA), PCNA (WL03213, 1:500, Wanleibio, China), CyclinE1 (WL01072, 1:1000, Wanleibio, China), CDK2 (WL01543, 1:500, Wanleibio, China), CyclinD1 (WL01435a, 1:400, Wanleibio, China), CDK6 (WL03460, 1:500, Wanleibio, China), Bcl-2 (WL01556, 1:1000, Wanleibio, China), EZH2 (WL00795, 1:1000, Wanleibio, China), p-ERK (WLP1512, 1:1000, Wanleibio, China), ERK (WL01864, 1:1000, Wanleibio, China), p-38 (WL00764, 1:1000, Wanleibio, China), and GAPDH (5174, 1:1000, Cell Signaling Technology, MA).

Techniques: Inhibition, Knockdown, CCK-8 Assay, Flow Cytometry, Cell Cycle Assay

Journal: Blood and Lymphatic Cancer: Targets and Therapy

Article Title: Crosstalk Between FOXN3 and E2F5 Reveals a Novel Tumor Suppressive Pathway in Acute Myeloid Leukemia via MAPK Signaling: Implications for Potential Future Targeted Therapy

doi: 10.2147/BLCTT.S571966

Figure Lengend Snippet: Enhancement of cell proliferation and cell cycle following E2F5 overexpression in AML cells. ( A ) CCK-8 assay evaluated the proliferation of Kasumi1 and THP1 cells following E2F5 overexpression. ( B ) Cell cycle analysis was conducted on Kasumi1 and THP1 cells with E2F5 overexpression using flow cytometry. Data are presented as mean ± SD of three independent experiments. Statistical significance is indicated as follows: ** P < 0.05, *** P < 0.01, and *** P < 0.0001.

Article Snippet: The primary antibodies used were as follows: FOXN3 (ab129453, 1:2500, Abcam, UK), E2F5 (A16042, 1:500, ABclonal, China), CDC42 (2466T, 1:1000, Cell Signaling Technology, MA), HDAC1 (34589T, 1:1000, Cell Signaling Technology, MA), PCNA (WL03213, 1:500, Wanleibio, China), CyclinE1 (WL01072, 1:1000, Wanleibio, China), CDK2 (WL01543, 1:500, Wanleibio, China), CyclinD1 (WL01435a, 1:400, Wanleibio, China), CDK6 (WL03460, 1:500, Wanleibio, China), Bcl-2 (WL01556, 1:1000, Wanleibio, China), EZH2 (WL00795, 1:1000, Wanleibio, China), p-ERK (WLP1512, 1:1000, Wanleibio, China), ERK (WL01864, 1:1000, Wanleibio, China), p-38 (WL00764, 1:1000, Wanleibio, China), and GAPDH (5174, 1:1000, Cell Signaling Technology, MA).

Techniques: Over Expression, CCK-8 Assay, Cell Cycle Assay, Flow Cytometry

Journal: Blood and Lymphatic Cancer: Targets and Therapy

Article Title: Crosstalk Between FOXN3 and E2F5 Reveals a Novel Tumor Suppressive Pathway in Acute Myeloid Leukemia via MAPK Signaling: Implications for Potential Future Targeted Therapy

doi: 10.2147/BLCTT.S571966

Figure Lengend Snippet: Partial rescue of proliferation inhibition and induction of apoptosis by E2F5 upregulation in AML cells overexpressing FOXN3. ( A ) Assessment of cell proliferation in leukemic cells co-transfected with lentiviruses for FOXN3 overexpression and E2F5 overexpression using the CCK-8 assay. ( B ) Flow cytometry analysis evaluated apoptosis in leukemic cells co-transfected with FOXN3 and E2F5 overexpressing lentiviruses. ( C ) Cell cycle analysis was performed on leukemic cells co-transfected with FOXN3 and E2F5 overexpressing lentiviruses using flow cytometry. Data are presented as mean ± SD of three independent experiments. Statistical significance is indicated as follows:* P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: The primary antibodies used were as follows: FOXN3 (ab129453, 1:2500, Abcam, UK), E2F5 (A16042, 1:500, ABclonal, China), CDC42 (2466T, 1:1000, Cell Signaling Technology, MA), HDAC1 (34589T, 1:1000, Cell Signaling Technology, MA), PCNA (WL03213, 1:500, Wanleibio, China), CyclinE1 (WL01072, 1:1000, Wanleibio, China), CDK2 (WL01543, 1:500, Wanleibio, China), CyclinD1 (WL01435a, 1:400, Wanleibio, China), CDK6 (WL03460, 1:500, Wanleibio, China), Bcl-2 (WL01556, 1:1000, Wanleibio, China), EZH2 (WL00795, 1:1000, Wanleibio, China), p-ERK (WLP1512, 1:1000, Wanleibio, China), ERK (WL01864, 1:1000, Wanleibio, China), p-38 (WL00764, 1:1000, Wanleibio, China), and GAPDH (5174, 1:1000, Cell Signaling Technology, MA).

Techniques: Inhibition, Transfection, Over Expression, CCK-8 Assay, Flow Cytometry, Cell Cycle Assay

Journal: Blood and Lymphatic Cancer: Targets and Therapy

Article Title: Crosstalk Between FOXN3 and E2F5 Reveals a Novel Tumor Suppressive Pathway in Acute Myeloid Leukemia via MAPK Signaling: Implications for Potential Future Targeted Therapy

doi: 10.2147/BLCTT.S571966

Figure Lengend Snippet: Regulation of EZH2 and MAPK pathways by the FOXN3-E2F5 axis in AML. ( A ) Volcano plots illustrating differentially expressed genes in THP1 cells with FOXN3 overexpression compared to control THP1 cells. ( B ) The top twenty significant KEGG pathways related to differentially expressed genes in THP1 cells with FOXN3 overexpression compared to control THP1 cells. The red box highlights the signal pathway that are in agreement with the functional results. ( C ) Western blot analysis showing the protein levels of EZH2, ERK1/2, and phosphorylated ERK1/2 (p-ERK1/2) in cells co-transfected with lentiviruses for FOXN3 and E2F5 overexpression. The experiments were conducted using three independent replicates and the data are presented as mean ± SD. Statistical significance is indicated as follows: * P < 0.05, and ** P < 0.01.

Article Snippet: The primary antibodies used were as follows: FOXN3 (ab129453, 1:2500, Abcam, UK), E2F5 (A16042, 1:500, ABclonal, China), CDC42 (2466T, 1:1000, Cell Signaling Technology, MA), HDAC1 (34589T, 1:1000, Cell Signaling Technology, MA), PCNA (WL03213, 1:500, Wanleibio, China), CyclinE1 (WL01072, 1:1000, Wanleibio, China), CDK2 (WL01543, 1:500, Wanleibio, China), CyclinD1 (WL01435a, 1:400, Wanleibio, China), CDK6 (WL03460, 1:500, Wanleibio, China), Bcl-2 (WL01556, 1:1000, Wanleibio, China), EZH2 (WL00795, 1:1000, Wanleibio, China), p-ERK (WLP1512, 1:1000, Wanleibio, China), ERK (WL01864, 1:1000, Wanleibio, China), p-38 (WL00764, 1:1000, Wanleibio, China), and GAPDH (5174, 1:1000, Cell Signaling Technology, MA).

Techniques: Over Expression, Control, Functional Assay, Western Blot, Transfection

Journal: Blood and Lymphatic Cancer: Targets and Therapy

Article Title: Crosstalk Between FOXN3 and E2F5 Reveals a Novel Tumor Suppressive Pathway in Acute Myeloid Leukemia via MAPK Signaling: Implications for Potential Future Targeted Therapy

doi: 10.2147/BLCTT.S571966

Figure Lengend Snippet: The mechanism of FOXN3-E2F5-EZH2-MAPK regulatory axis in AML.

Article Snippet: The primary antibodies used were as follows: FOXN3 (ab129453, 1:2500, Abcam, UK), E2F5 (A16042, 1:500, ABclonal, China), CDC42 (2466T, 1:1000, Cell Signaling Technology, MA), HDAC1 (34589T, 1:1000, Cell Signaling Technology, MA), PCNA (WL03213, 1:500, Wanleibio, China), CyclinE1 (WL01072, 1:1000, Wanleibio, China), CDK2 (WL01543, 1:500, Wanleibio, China), CyclinD1 (WL01435a, 1:400, Wanleibio, China), CDK6 (WL03460, 1:500, Wanleibio, China), Bcl-2 (WL01556, 1:1000, Wanleibio, China), EZH2 (WL00795, 1:1000, Wanleibio, China), p-ERK (WLP1512, 1:1000, Wanleibio, China), ERK (WL01864, 1:1000, Wanleibio, China), p-38 (WL00764, 1:1000, Wanleibio, China), and GAPDH (5174, 1:1000, Cell Signaling Technology, MA).

Techniques:

Journal: Translational Andrology and Urology

Article Title: Fu Zheng Xiao Yu San Jie Decoction affects the proliferation of renal cell carcinoma via regulating E2F5 gene

doi: 10.21037/tau-2025-aw-739

Figure Lengend Snippet: Potential target genes of FZXYJD in E2F family were predicted. (A) E2F5 has low expression in ccRCC. (B) Patients had poorer survival with low expression of E2F5 in ccRCC. (C-F) FZXYJD can increase the expression level of E2F5 in ccRCC cell line. *, P<0.05; **, P<0.01; ***, P<0.001. Control, cultured in SD rat serum. LD, cultured in FZXYJD lower dose group serum. MD, cultured in FZXYJD medium dose group serum. HD, cultured in FZXYJD high-dose group serum. ccRCC, clear cell renal cell carcinoma; FZXYSJD, Fu Zheng Xiao Yu San Jie Decoction; HD, high-dose; HR, hazard ratio; KIRC, kidney renal clear cell carcinoma; LD, low-dose; MD, medium-dose; N, normal; SD, Sprague-Dawley; T, tumor; TPM, transcripts per million.

Article Snippet: It was then incubated overnight at 4 °C with the primary antibodies: E2F5 (1:1,000, Catalog: D197169, BBI, Shanghai, China) and β-Actin (1:1,000, Catalog: ab8227, Abcam, Shanghai, China).

Techniques: Expressing, Control, Cell Culture

Journal: Translational Andrology and Urology

Article Title: Fu Zheng Xiao Yu San Jie Decoction affects the proliferation of renal cell carcinoma via regulating E2F5 gene

doi: 10.21037/tau-2025-aw-739

Figure Lengend Snippet: E2F5 was involved in FZXYJD suppression of ccRCC cells proliferation. (A) Western blot and RT-qPCR were performed to analyze E2F5 expression in si E2F5 -transfected ccRCC cell line. β-actin was used as an internal quantitative control. (B) CCK-8 assay. (C) Cell colony formation assay [IPHONE X PRO MAX] (magnification: 1×; crystal violet stained). *, P<0.05; **, P<0.01; ***, P<0.001. CCK-8, Cell Counting Kit-8; ccRCC, clear cell renal cell carcinoma; FZXYSJD, Fu Zheng Xiao Yu San Jie Decoction; NC, negative control; OD, optical density; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; si E2F5 , small interfering RNA targeting E2F5 .

Article Snippet: It was then incubated overnight at 4 °C with the primary antibodies: E2F5 (1:1,000, Catalog: D197169, BBI, Shanghai, China) and β-Actin (1:1,000, Catalog: ab8227, Abcam, Shanghai, China).

Techniques: Western Blot, Quantitative RT-PCR, Expressing, Transfection, Control, CCK-8 Assay, Colony Assay, Staining, Cell Counting, Negative Control, Reverse Transcription, Real-time Polymerase Chain Reaction, Small Interfering RNA

Journal: Translational Andrology and Urology

Article Title: Fu Zheng Xiao Yu San Jie Decoction affects the proliferation of renal cell carcinoma via regulating E2F5 gene

doi: 10.21037/tau-2025-aw-739

Figure Lengend Snippet: Three-dimensional and two-dimensional docking patterns and interactions of FZXYSJD identified components with the E2F5 protein. FZXYSJD, Fu Zheng Xiao Yu San Jie Decoction.

Article Snippet: It was then incubated overnight at 4 °C with the primary antibodies: E2F5 (1:1,000, Catalog: D197169, BBI, Shanghai, China) and β-Actin (1:1,000, Catalog: ab8227, Abcam, Shanghai, China).

Techniques: