pfkfb4 antibody  (GeneTex)

Journal: Scientific Reports

Article Title: Differential roles of highly expressed PFKFB4 in colon adenocarcinoma patients

doi: 10.1038/s41598-023-43619-4

Figure Lengend Snippet: PFKFB4 expression and clinicopathological characteristics of patients with COAD.

Article Snippet: Next, the sections were incubated with anti-PFKFB4 antibody (1:100; Cat. No. ab137785; Abcam) at 37 °C for 1 h, followed by overnight incubation in a humid environment at 4 °C.

Techniques: Expressing

Journal: Scientific Reports

Article Title: Differential roles of highly expressed PFKFB4 in colon adenocarcinoma patients

doi: 10.1038/s41598-023-43619-4

Figure Lengend Snippet: Representative immunohistochemical images of PFKFB4 expression in COAD tissues. This figure presents representative immunohistochemical images depicting the expression of PFKFB4 in COAD tissues. The images demonstrate the staining intensity at different magnifications: Negative staining (corresponding normal (control) tissue) at magnifications ( a ) × 100 and ( e ) × 400. Weak staining at magnifications ( b ) × 100 and ( f) × 400. Moderate staining at magnifications ( c ) × 100 and ( g ) × 400. Strong positive staining at magnifications ( d ) × 100 and ( h ) × 400.

Article Snippet: Next, the sections were incubated with anti-PFKFB4 antibody (1:100; Cat. No. ab137785; Abcam) at 37 °C for 1 h, followed by overnight incubation in a humid environment at 4 °C.

Techniques: Immunohistochemistry, Expressing, Staining, Negative Staining

Journal: Scientific Reports

Article Title: Differential roles of highly expressed PFKFB4 in colon adenocarcinoma patients

doi: 10.1038/s41598-023-43619-4

Figure Lengend Snippet: Elevated PFKFB4 expression in cancerous tissues compared with adjacent normal tissues. ( a ) The figure presents a box plot displaying the distribution of PFKFB4 staining scores in adjacent normal tissues (n = 15) and COAD tissues (n = 79). The box plot shows the median with the interquartile range. Statistical analysis was performed using the unpaired Student’s t-test to calculate the p -value. ( b ) A comparison of PFKFB4 expression between 15 pairs of matched neighboring normal tissues and COAD tissues. The analysis utilized the paired Student’s t-test to determine the p -value.

Article Snippet: Next, the sections were incubated with anti-PFKFB4 antibody (1:100; Cat. No. ab137785; Abcam) at 37 °C for 1 h, followed by overnight incubation in a humid environment at 4 °C.

Techniques: Expressing, Staining, Comparison

Journal: Scientific Reports

Article Title: Differential roles of highly expressed PFKFB4 in colon adenocarcinoma patients

doi: 10.1038/s41598-023-43619-4

Figure Lengend Snippet: PFKFB4 expression in various cancer types. This figure depicts the expression levels of PFKFB4 in different types of cancer. The data for PFKFB4 expression were analyzed using the TIMER2.0 database, encompassing multiple cancer types. The significance levels for the analysis are represented as follows: * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: Next, the sections were incubated with anti-PFKFB4 antibody (1:100; Cat. No. ab137785; Abcam) at 37 °C for 1 h, followed by overnight incubation in a humid environment at 4 °C.

Techniques: Expressing

Journal: Scientific Reports

Article Title: Differential roles of highly expressed PFKFB4 in colon adenocarcinoma patients

doi: 10.1038/s41598-023-43619-4

Figure Lengend Snippet: Relationship between high expression of PFKFB4 and RFS, OS, and PPS in COAD patients. This figure illustrates the association between high expression of PFKFB4 and three clinical outcomes in COAD patients: Relapse-Free Survival (RFS), Overall Survival (OS), and Post-Progression Survival (PPS). The analysis utilized Kaplan–Meier survival analysis on data obtained from the Kaplan–Meier plotter database. A subsequent log-rank test was performed to assess the statistical significance of the results.

Article Snippet: Next, the sections were incubated with anti-PFKFB4 antibody (1:100; Cat. No. ab137785; Abcam) at 37 °C for 1 h, followed by overnight incubation in a humid environment at 4 °C.

Techniques: Expressing

Journal: Scientific Reports

Article Title: Differential roles of highly expressed PFKFB4 in colon adenocarcinoma patients

doi: 10.1038/s41598-023-43619-4

Figure Lengend Snippet: Histogram of functional analysis of co-expressed genes of PFKFB4 in COAD. In this figure, we present the functional analysis of co-expressed genes associated with PFKFB4 in COAD. A total of 10,189 PFKFB4-related genes in COAD were identified through co-expression analysis. These co-expressed genes were further subjected to classification and functional analyses using the following enrichment databases: ( a ) Gene Ontology (GO) enrichment analysis; ( b ) Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis; ( c ) Reactome enrichment analysis. The CAMOIP database was used to perform these enrichment analyses.

Article Snippet: Next, the sections were incubated with anti-PFKFB4 antibody (1:100; Cat. No. ab137785; Abcam) at 37 °C for 1 h, followed by overnight incubation in a humid environment at 4 °C.

Techniques: Functional Assay, Expressing

Journal: Scientific Reports

Article Title: Differential roles of highly expressed PFKFB4 in colon adenocarcinoma patients

doi: 10.1038/s41598-023-43619-4

Figure Lengend Snippet: Bubble diagrams of functional analysis of co-expressed genes of PFKFB4 in COAD. This figure presents the functional analysis of co-expressed genes associated with PFKFB4 in COAD. A total of 10,189 PFKFB4-related genes in COAD were identified through co-expression analysis. The top 20 enriched pathways from the co-expressed genes of PFKFB4 were analyzed using the following enrichment databases: ( a ) Gene Ontology (GO) enrichment analysis; ( b ) Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis; ( c ) Reactome enrichment analysis. The enrichment analyses were performed using the CAMOIP database.

Article Snippet: Next, the sections were incubated with anti-PFKFB4 antibody (1:100; Cat. No. ab137785; Abcam) at 37 °C for 1 h, followed by overnight incubation in a humid environment at 4 °C.

Techniques: Functional Assay, Expressing

Journal: Scientific Reports

Article Title: Differential roles of highly expressed PFKFB4 in colon adenocarcinoma patients

doi: 10.1038/s41598-023-43619-4

Figure Lengend Snippet: Correlation of PFKFB4 expression with immune infiltrates in COAD. This figure demonstrates the correlation between PFKFB4 expression and immune infiltrates in COAD. The immune cell scores estimated by different methods were compared between different groups using the Mann–Whitney U-test based on the CAMOIP database. Comparison of immune cell scores estimated by ( a ) CIBERSORT, ( b ) EPIC, ( c ) IPS, ( d ) MCPcounter, and ( e ) quanTIseq, respectively. The significance levels for the comparisons are indicated as follows: * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Next, the sections were incubated with anti-PFKFB4 antibody (1:100; Cat. No. ab137785; Abcam) at 37 °C for 1 h, followed by overnight incubation in a humid environment at 4 °C.

Techniques: Expressing, MANN-WHITNEY, Comparison

Journal: Cell Death & Disease

Article Title: Hypoxia-mediated promotion of glucose metabolism in non-small cell lung cancer correlates with activation of the EZH2/FBXL7/PFKFB4 axis

doi: 10.1038/s41419-023-05795-z

Figure Lengend Snippet: A PFKFB4 expression was elevated in NSCLC tissue samples, as analyzed by UALCAN database. LUSC: lung squamous cell carcinoma; LUAD: lung adenocarcinoma. B Kaplan–Meier analysis showing the negative correlation between PFKFB4 and the overall survival rate of NSCLC patients. Red: high expression of PFKFB4; black: low expression of PFKFB4. C Co-IP analysis of the interaction of endogenous FBXL7 with PFKFB4 in 293T cells. D Co-IP analysis of the interaction of exogenous FBXL7 with PFKFB4 in 293T cells. E GST-pull down assay analysis of the direct interaction between FBXL7 and PFKFB4 in 293T cells. F PFKFB4 mRNA expression in A549 cells wasn’t changed in response to oe-FBXL7, as determined by RT-qPCR. G Western blot analysis showing inhibited PFKFB4 protein expression in A549 cells in response to oe-FBXL7. H Western blot analysis showing enhanced degradation of PFKFB4 protein in 293T cells in response to oe-FBXL7 and CHX. I Ubiquitination level of PFKFB4 protein was reduced in 293T cells in the presence of si-FBXL7, as determined by IP assay. J Ubiquitination level of PFKFB4 protein was elevated in 293T cells in the presence of oe-FBXL7, as determined by IP assay. ** p < 0.01, *** p < 0.001, **** p < 0.0001. ns p > 0.05. The cell experiment was repeated three times independently.

Article Snippet: The immunohistochemical staining included primary antibodies against FBXL7 (sc-374319, 1:100, Santa Cruz), Ki-67 (ab15580, 1:100, Abcam), EZH2 (5246, 1:100, CST), and PFKFB4 (ab137785, 1:100, Abcam).

Techniques: Expressing, Co-Immunoprecipitation Assay, Pull Down Assay, Quantitative RT-PCR, Western Blot

Journal: Cell Death & Disease

Article Title: Hypoxia-mediated promotion of glucose metabolism in non-small cell lung cancer correlates with activation of the EZH2/FBXL7/PFKFB4 axis

doi: 10.1038/s41419-023-05795-z

Figure Lengend Snippet: A Western blot analysis of FBXL7 and PFKFB4 proteins in A549 cells in response to oe-FBXL7, oe-FBXL7 + oe-PFKFB4, or oe-FBXL7 + oe-PFKFB4 + 2-DG. B Glucose uptake measurement in response to oe-FBXL7, oe-FBXL7 + oe-PFKFB4, or oe-FBXL7 + oe-PFKFB4 + 2-DG. C Pyruvate level measurement in response to oe-FBXL7, oe-FBXL7 + oe-PFKFB4, or oe-FBXL7 + oe-PFKFB4 + 2-DG. D Lactate production measurement in response to oe-FBXL7, oe-FBXL7 + oe-PFKFB4, or oe-FBXL7 + oe-PFKFB4 + 2-DG. E ATP level measurement in response to oe-FBXL7, oe-FBXL7 + oe-PFKFB4, or oe-FBXL7 + oe-PFKFB4 + 2-DG. F ROS content measurement in A549 cells in response to oe-FBXL7, oe-FBXL7 + oe-PFKFB4, or oe-FBXL7 + oe-PFKFB4 + 2-DG. G ROS content measurement in mitochondrion in response to oe-FBXL7, oe-FBXL7 + oe-PFKFB4, or oe-FBXL7 + oe-PFKFB4 + 2-DG. H Glycolysis of A549 cells in response to oe-FBXL7, oe-FBXL7 + oe-PFKFB4, or oe-FBXL7 + oe-PFKFB4 + 2-DG as detected by ECAR assay ( # p < 0.05 vs. the oe-FBXL7 group; @ p < 0.05 vs. the oe-FBXL7 + oe-PFKFB4 group). I Viability of A549 cells in response to oe-FBXL7, oe-FBXL7 + oe-PFKFB4, or oe-FBXL7 + oe-PFKFB4 + 2-DG, as analyzed by CCK-8 assay. J Migration and invasion of A549 cells in response to oe-FBXL7, oe-FBXL7 + oe-PFKFB4, or oe-FBXL7 + oe-PFKFB4 + 2-DG, as analyzed by Transwell assay. K Apoptosis of A549 cells in response to oe-FBXL7, oe-FBXL7 + oe-PFKFB4, or oe-FBXL7 + oe-PFKFB4 + 2-DG, as analyzed by flow cytometry. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. ns p > 0.05. The cell experiment was repeated three times independently.

Article Snippet: The immunohistochemical staining included primary antibodies against FBXL7 (sc-374319, 1:100, Santa Cruz), Ki-67 (ab15580, 1:100, Abcam), EZH2 (5246, 1:100, CST), and PFKFB4 (ab137785, 1:100, Abcam).

Techniques: Western Blot, ECAR Assay, CCK-8 Assay, Migration, Transwell Assay, Flow Cytometry

Journal: Cell Death & Disease

Article Title: Hypoxia-mediated promotion of glucose metabolism in non-small cell lung cancer correlates with activation of the EZH2/FBXL7/PFKFB4 axis

doi: 10.1038/s41419-023-05795-z

Figure Lengend Snippet: A Western blot analysis showing elevated expression of EZH2 and HIF-1α proteins in hypoxic A549 cells. B Verification of the binding of HIF-1α to the EZH2 promoter region, as evaluated by dual-luciferase reporter assay. C Enrichment of HIF-1α in the promoter region of EZH2, as observed by ChIP assay. D mRNA expression of EZH2, FBXL7, and PFKFB4 in normoxic or hypoxic A549 cells in the presence of si-EZH2. E Western blot analysis showing the expression of EZH2, FBXL7, and PFKFB4 proteins in normoxic or hypoxic A549 cells in the presence of si-EZH2. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. ns p > 0.05. The cell experiment was repeated three times independently.

Article Snippet: The immunohistochemical staining included primary antibodies against FBXL7 (sc-374319, 1:100, Santa Cruz), Ki-67 (ab15580, 1:100, Abcam), EZH2 (5246, 1:100, CST), and PFKFB4 (ab137785, 1:100, Abcam).

Techniques: Western Blot, Expressing, Binding Assay, Luciferase, Reporter Assay

Journal: Cell Death & Disease

Article Title: Hypoxia-mediated promotion of glucose metabolism in non-small cell lung cancer correlates with activation of the EZH2/FBXL7/PFKFB4 axis

doi: 10.1038/s41419-023-05795-z

Figure Lengend Snippet: A The mRNA expression of EZH2, FBXL7 and PFKFB4 in hypoxia-exposed A549 cells in response to si-EZH2, si-EZH2 + si-FBXL7 or si-EZH2 + si-FBXL7 + 2-DG. B Western blot analysis showing the expression of EZH2, FBXL7, and PFKFB4 proteins in hypoxia-exposed A549 cells in response to si-EZH2, si-EZH2 + si-FBXL7 or si-EZH2 + si-FBXL7 + 2-DG. C Glucose uptake measurement in response to si-EZH2, si-EZH2 + si-FBXL7, or si-EZH2 + si-FBXL7 + 2-DG. D Pyruvate level measurement in response to si-EZH2, si-EZH2 + si-FBXL7, or si-EZH2 + si-FBXL7 + 2-DG. E Lactate production measurement in response to si-EZH2, si-EZH2 + si-FBXL7, or si-EZH2 + si-FBXL7 + 2-DG. F ATP level measurement in response to si-EZH2, si-EZH2 + si-FBXL7, or si-EZH2 + si-FBXL7 + 2-DG. G ROS content measurement in A549 cells in response to si-EZH2, si-EZH2 + si-FBXL7 or si-EZH2 + si-FBXL7 + 2-DG. H ROS content measurement in mitochondrion in response to si-EZH2, si-EZH2 + si-FBXL7, or si-EZH2 + si-FBXL7 + 2-DG. I Glycolysis of A549 cells in response to si-EZH2, si-EZH2 + si-FBXL7, or si-EZH2 + si-FBXL7 + 2-DG, as detected by ECAR assay ( # p < 0.05 vs. the Hypoxia + si-EZH2 group; @ p < 0.05 vs. the Hypoxia + si-NC + si-FBXL7 group). J Viability of A549 cells in response to si-EZH2, si-EZH2 + si-FBXL7 or si-EZH2 + si-FBXL7 + 2-DG, as analyzed by CCK-8 assay. K Migration and invasion of A549 cells, as analyzed by Transwell assay. L Apoptosis of A549 cells in response to si-EZH2, si-EZH2 + si-FBXL7, or si-EZH2 + si-FBXL7 + 2-DG, as analyzed by flow cytometry. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. ns p > 0.05. The cell experiment was repeated three times independently.

Article Snippet: The immunohistochemical staining included primary antibodies against FBXL7 (sc-374319, 1:100, Santa Cruz), Ki-67 (ab15580, 1:100, Abcam), EZH2 (5246, 1:100, CST), and PFKFB4 (ab137785, 1:100, Abcam).

Techniques: Expressing, Western Blot, ECAR Assay, CCK-8 Assay, Migration, Transwell Assay, Flow Cytometry

Journal: Cell Death & Disease

Article Title: Hypoxia-mediated promotion of glucose metabolism in non-small cell lung cancer correlates with activation of the EZH2/FBXL7/PFKFB4 axis

doi: 10.1038/s41419-023-05795-z

Figure Lengend Snippet: A Quantitative analysis for tumor volume of nude mice subcutaneously inoculated with cells transduced with sh-EZH2 alone or combined with oe-PFKFB4. B Quantitative analysis for tumor weight of nude mice subcutaneously inoculated with cells transduced with sh-EZH2 alone or combined with oe-PFKFB4. C HE staining analysis of the tumor tissues of nude mice subcutaneously inoculated with cells transduced with sh-EZH2 alone or combined with oe-PFKFB4. D Immunohistochemical staining analysis of Ki-67, EZH2, PFKFB4, and FBXL7 proteins in the tumor tissues of nude mice subcutaneously inoculated with cells transduced with sh-EZH2 alone or combined with oe-PFKFB4. E TUNEL-positive cells in the tumor tissues of nude mice subcutaneously inoculated with cells transduced with sh-EZH2 alone or combined with oe-PFKFB4. *** p < 0.001, **** p < 0.0001. ns p > 0.05. n = 6 for mice in each group.

Article Snippet: The immunohistochemical staining included primary antibodies against FBXL7 (sc-374319, 1:100, Santa Cruz), Ki-67 (ab15580, 1:100, Abcam), EZH2 (5246, 1:100, CST), and PFKFB4 (ab137785, 1:100, Abcam).

Techniques: Transduction, Staining, Immunohistochemical staining, TUNEL Assay

Journal: Cell Death & Disease

Article Title: Hypoxia-mediated promotion of glucose metabolism in non-small cell lung cancer correlates with activation of the EZH2/FBXL7/PFKFB4 axis

doi: 10.1038/s41419-023-05795-z

Figure Lengend Snippet: Hypoxia induces the expression of HIF-1α in NSCLC cells. HIF-1α elevates expression of EZH2 which in turn reduces expression of FBXL7, and inhibits the ubiquitination of PFKFB4 by FBXL7, resulting in upregulation of PFKFB4 protein expression. By this mechanism, NSCLC cell glycolysis and malignant phenotypes are accelerated while cell apoptosis was decelerated.

Article Snippet: The immunohistochemical staining included primary antibodies against FBXL7 (sc-374319, 1:100, Santa Cruz), Ki-67 (ab15580, 1:100, Abcam), EZH2 (5246, 1:100, CST), and PFKFB4 (ab137785, 1:100, Abcam).

Techniques: Expressing