HBV Upregulates CtBP2 Expression via the X Gene

Figures

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  Figure 1. CtBP2 expression in HBV-related HCC tissues. (a) Total RNA was extracted using TRIzol from thirty-five pairs of HBV-related HCC tissues and paired paracancerous liver tissues, and CtBP2 mRNA expression was detected by RT-qPCR. (b) Total protein was extracted from thirty-five pairs of HBV-related HCC tissues and paired paracancerous liver tissues, and CtBP2 protein expression was detected by western blotting.∗p < 0.05.

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  Figure 2. Effect of HBV on CtBP2 expression in HepG2 cells. (a) TRIzol was used to extract the total RNA from HepG2.2.15 cells integrated with the HBV genome and the control HepG2 cells, and mRNA expression was detected by RT-qPCR. (b) Total protein was extracted from the HepG2.2.15 cells and the control HepG2 cells, and protein expression was detected using western blotting; (c) HepG2 cells were transfected with 0.6 μg of HBV1.3 and the control plasmid pBlue-ks, and CtBP2 mRNA expression was detected by RT-qPCR after 48 h; (d) HepG2 cells were transfected with 0.6 μg of HBV1.3 and the control plasmid pBlue-ks, and CtBP2 protein expression was detected by western blotting after 48 h.

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  Figure 3. Effect of the HBV X gene on CtBP2 promoter activity. (a) Luciferase activity was measured 48 h after cotransfection of 0.6 μg of the eukaryotic expression plasmid containing each gene of the HBV genome (pCMV-S, pCMV-E, pCMV-C, pCMV-X, and pCMV-P) and 0.2 μg of the CtBP2 gene promoter pGL3-CtBP2 into HepG2 cells using pCMV-tag2B as the blank control. Each experiment was repeated three times. (b) Different doses of pCMV-X (0 μg, 0.2 μg, 0.4 μg, 0.6 μg, and 0.8 μg) and pGL3-CtBP2 were cotransfected into HepG2 cells, and the luciferase activity was detected after 48 h. ∗p < 0.05.

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  Figure 4. Effect of the HBV X gene on CtBP2 mRNA and protein expression. (a) CtBP2 mRNA expression was detected by RT-qPCR 48 h after transfection of different doses of the X gene eukaryotic expression plasmid pCMV-X (0 μg, 0.5 μg, 1 μg, and 2 μg) into HepG2 cells. (b) CtBP2 protein expression was detected by western blotting 48 h after transfection of different doses of the X gene eukaryotic expression plasmid pCMV-X (0 μg, 0.5 μg, 1 μg, and 2 μg) into HepG2 cells. (c) Correlation between HBx and CtBP2 expression in HBV-related HCC tissues.