Figure 4 | Scientific Reports

Figure 4

From: Epigenetically regulated miR-449a enhances hepatitis B virus replication by targeting cAMP-responsive element binding protein 5 and modulating hepatocytes phenotype

Figure 4

CREB5 is directly targeted by miR-449a and is involved in the control of HBV replication.

(A) HepG2.2.15 cells were transfected with 20 nM of different siRNAs targeting MMAB, GAL, DDI2, TMEM194B, CREB5, MCM10, SASS6, MYBL1, ZMAT1 and APOBEC3B for 4 days. HBV replication was detected by Southern blotting. (B) Western blot analysis of CREB5 and FXRα protein expression in HepG2.2.15 cells transfected with miR-449a or siRNA specific for CREB5 at 20 nM for 3 days. (C) Wildtype and mutated pmiR-CREB5-3UTR luciferase reporters (100 ng each) were co-transfected with 20 nM of miR-449a in Huh7 cells, luciferase activity was assayed at 48 h. The fold change of luciferase expression expressed as the ratio of miR-449a- to miR-con-transfected samples. (D) The CREB5 expression vector pcDNA3.1/V5-CREB5 was co-transfected in Huh7 cells with pSM2 at the indicated concentrations for 4 days. HBV replication was detected by Southern blotting and CREB5 expression was determined by V5-tag western blotting. The levels of secreted HBsAg and HBeAg in culture media were measured by the CMIA test. *P < 0.05.

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