Fig 1: Expression and methylation of E2F2 in GC tissues and normal tissues as revealed by bioinformatic analysis. (A) E2F2 expression levels in different tumor types from the TCGA database were detected by TIMER (*P < 0.05, **P < 0.01, ***P < 0.001). (B and C) E2F2 mRNA is highly expressed in LGG tissues in GEPIA (B) dataset and Oncomine (C) dataset. (D) Kaplan-Meier analysis of survival rates of GC patients with high E2F2 expression and GC patients with low E2F2 expression.
Fig 2: Expression of E2F2 was related to a panel of gene markers of immune cells, including M1 cells (A), M2 cells (B), Th1 cells (C), Th2 cells (D), Treg cells (E) and PD-1/PD-L1 (F). (G) E2F2 CNV affects the infiltrating levels of B cells, CD8+ T cells, CD4+ T cells, macrophages, neutrophils and dendritic cells in GC.
Fig 3: E2F2 mRNA expression in subgroups of patients with GC, stratified based on gender, age and other criteria (UALCAN). (A) Boxplot shows the relative expression of E2F2 in normal and STAD samples. (B) Boxplot shows the relative expression of E2F2 in normal individuals of any age and in STAD patients aged 21-40, 41-60, 61-80, or 81-100 years. (C) Boxplot shows the relative expression of E2F2 in normal individuals of either gender and in male or female STAD patients. (D) Boxplot shows the relative expression of E2F2 in normal individuals and in STAD patients with stage 1, 2, 3 or 4 disease. (E) Boxplot shows the relative expression of E2F2 in normal individuals and in STAD patients with grade 1, 2, 3 or 4 tumors. (F) Boxplot shows the relative expression of E2F2 based on TP53 mutation status. The central mark is the median; the edges of the box are the 25th and 75th percentiles. A t-test was used to estimate the significance of differences in gene expression levels between groups. *P < 0.05; **P < 0.01; ***P < 0.001.
Fig 4: Translational study of E2F2 degrader in hepatocellular carcinoma growth in vivo. a, Schematic diagram of the administration procedure of bufalin in the established H22 tumor-bearing mice. Cyclophosphamide (CTX) was used as a positive control. b, The dynamic change of tumor volume in subcutaneous models were displayed. c, Bufalin decreased tumor volume in ICR mice transplanted with H22 cells (n = 6). d, Bufalin obviously reduced tumor weight in H22 xenograft mice compared with vehicle group (n = 6). e, bufalin treatment had no significant effect on the body weight of H22-xenograft mice. f, bufalin treatment did not significantly change the weight of heart, liver, spleen, lung, and kidney in H22-xenograft mice (n = 6). g, Representative histological analysis of tumor specimen stained by H&E (Scale bars = 100 µm). h, Representative images of IHC staining for E2F2 in tumor tissues. Data were presented as mean ± SD. ***P < 0.001, by one-way ANOVA. N.S. not significant.
Fig 5: Protein-protein interaction (PPI) network construction and gene enrichment analyses. (A) Network of E2F2 and its 50 frequently altered neighbor genes was constructed. (B) Hub genes were screened from the PPI network using the Closeness, Degree and MCC methods. (C) Functional enrichment histogram of important modules. Each biological process, cellular component and molecular function category is represented by a red, blue and green bar, respectively. The height represents the number of IDs in the user list and in the category. (D) Pathways enrichment map of E2F2 and its 50 frequently altered neighbor genes. The top 20 terms with the largest number of enriched genes were selected.
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