Fig 1: E2F1 regulates aerobic glycolysis gene expression of ENO2. (A) Western blot assays revealed the protein levels of E2F1, ENO2, PFKM and TPI11 in RDES cells treated with stably transfected as indicated. (B-E) Protein expression was quantified by densitometry using Image-Pro Plus with β-actin as the loading control and statistics were performed in GraphPad Prism 9. (F-H) The positive gene expression correlation between E2F1 and (F) ENO2, (G) PFKM and (H) TPI1. Student's t-test compared the difference in Mock vs. E2F1. One-way ANOVA followed by post-hoc Bonferroni's test was used to compare the difference in sh-Scb vs. sh-E2F1 #1 or sh-E2F1 #2. ***P<0.001 vs. Mock or sh-Scb; ns, no significance; E2F1, E2F transcription factor 1; sh, short hairpin.
Fig 2: E2F1 regulates aerobic glycolysis. The levels of (A) glucose uptake, (B) lactate production and (C) cellular ATP levels were detected in RDES cells upon the transfection of the indicated plasmids. Reverse transcription-quantitative PCR (normalized to β-actin, n=3) revealing the transcript levels of (D) E2F1, (E) ENO2, (F) PFKM and (G) TPI11 in RDES cells treated with stably transfected as indicated. Student's t-test compared the difference in Mock vs. E2F1. One-way ANOVA followed by post-hoc Bonferroni's test was used to compare the difference in sh-Scb vs. sh-E2F1 #1 or sh-E2F1 #2. ***P<0.001 vs. Mock or sh-Scb; ns, no significance; E2F1, E2F transcription factor 1; sh, short hairpin.
Fig 3: Expression of E2F1 and glycolytic genes in tumor tissues. (A) The relative E2F1 levels in EWS (GSE17679) with the mortality and progression. (B) Kaplan-Meier curves showing the overall survival in EWS (GSE17679) with high or low expression of E2F1 (cutoff values=5.998). (C) Kaplan-Meier curves showing the overall survival in EWS (GSE17679) with high or low expression of ENO2, PFKM, TPI1, and E2F1 (cutoff values=7.347, 9.177, 10.900 and 5.998). Student's t-test compared the difference in panel A. Log-rank test for survival comparison in panels B and C. E2F1, E2F transcription factor 1; EWS, Ewing sarcoma.
Fig 4: E2F1 involvement in aerobic glycolysis in EWS. (A) (left and right panel) Venn diagrams showing different expressed glycolytic genes and transcription factors (P<0.05) in EWS (GSE17679) associated with the status of mortality and progression, respectively; (middle panel) overlapping analysis with 15 transcription factors regulating three glycolytic genes analyzed by ChIP-X. (B) The relative ENO2 (upper panel), PFKM (middle panel) and TPI1 (lower panel) levels in EWS (GSE17679) with the mortality and progression. (C) Kaplan-Meier curves showing the overall survival in EWS (GSE17679) with high or low levels of ENO2, PFKM and TPI1 (cutoff values=7.474, 9.177 and 10.900). Student's t-test compared the difference in panel B. Log-rank test for survival comparison in panel C. E2F1, E2F transcription factor 1; EWS, Ewing sarcoma.
Fig 5: Immunoblotting analysis for validation of the differential expression of 11 proteins in ARC-KO cells. Immunoblotting (A) and quantification of protein expression (B) confirmed the differential expression of 11 proteins (ARC, HSPA1A, ENO1, ENO2, VCP, TUBB, FLNC, HMGCS1, ALDH1B1, FSCN1, and HINT2) in ARC-KO and ARC-WT HEK293 cells. GAPDH was used as the loading control. Data are expressed as fold changes relative to ARC-WT ± SD (* p < 0.05; n = 3).
Supplier Page from ABclonal Technology for ENO2 Rabbit pAb
Trial Size: 20 ul