Fig 1: Recruitment of components of the PRC2 and PRC1 complexes onto the KSHV genome during de novo infection.(A) Time-course ChIP analysis of the binding of EZH2 (PRC2), RING1B and RYBP (PRC1) onto KSHV promoters at 4, 24, and 72 hpi and during latency. (B) Genome-wide recruitment of EZH2 and RING1B to the KSHV genome at 4 and 72 hpi. The Pearson correlation between the binding of EZH2 and RING1B is 0.6 at 72 hpi. Labels are the same as in Figure 3.
Fig 2: Simultaneously knockdown of TXNIP can partially rescue the phenotype in RNF2 single knockdown DU145 cellsA. RT-qPCR analysis to show the efficient inhibition of RNF2 and TXNIP mRNA level in DU145 cells infected with shRNA expressing lentivirus. B. Western blot analysis to show the efficient knockdown of RNF2 and TXNIP expression in DU145 cells infected with shRNA expressing lentivirus. C. Cell cycle analysis in RNF2 and/or TXNIP knockdown and control DU145 cells. Data are shown as the mean ± S.D. from three independent experiments. D. Apoptosis analysis in RNF2 and/or TXNIP knockdown and control DU145 cells. Data are representative of three independent experiments. E. MTT assay to show the cell proliferation in RNF2 and/or TXNIP knockdown and control DU145 cells. Data are shown as the mean ± S.D. from three independent experiments. *p<0.05 versus shScr, Δp<0.05 versus shRNF2 #1.
Fig 3: PCa tumor tissues showed higher RNF2 expression than BPH tissuesA. Representative IHC staining to show RNF2 expression in PCa and BPH tissues. The brown color indicates the positive expression of RNF2. Right pictures are the magnified view of the yellow box in the left. Scale bar: 200 μm. B. Statistical analysis of H-score of RNF2 expression in PCa and BPH tissues. C. Statistical analysis of H-score of RNF2 expression in low and high grade PCa.
Fig 4: The downregulation of RNF2 induced EGR1 expression. (A) Heat map of some of the most differentially expressed genes in shRNF2-treated HCT116 cells, determined through microarray analyses (red: upregulated; green: downregulated). (B) RT-PCR analysis showing the increased EGR1 mRNA levels in RNF2-knockdown HCT116 cells. (C) Western blot showing the increased EGR1 protein levels in RNF2-knockdown HCT116 cells.
Fig 5: Overall survival curve of patients with esophageal squamous cell carcinoma in the RNF2+P-AKTpositive expression group compared with other group. RNF2, Ring finger protein 2; P-AKT, phosphor-protein kinase B; Other, including patients with RNF2-negative/P-AKT-negative, RNF2-positive/P-AKT-negative, RNF2-negative/P-AKT-positive.
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