SENP3 silencing suppresses HBV gene expression.

(A) RT-qPCR measurements of HBV transcripts amplified by primers HBV-PC and HBV-X in HepG2.215-control cells and HepG2.215-SENP3 K.D. cells. Beta-actin was used as internal control; the data were expressed as mean±SD (n = 3). The statistical significance was assessed by Students’ unpaired t-test. (B) RT-qPCR measurement of HBV transcripts amplified by primer pair HBV-X in HepG2.215-SENP3 K.D. cells with transient transfection of pcDNA3 plasmid (Mock) and RGS-SENP3 plasmid (SENP3 O.E.). Beta-actin was used as internal control; the data were expressed as mean±SD (n = 3). The statistical significance was assessed by Students’ unpaired t-test. (C) Immunoblotting of HBx and HBc in HepG2.215 control cells and HepG2.215 SENP3 K.D. cells. (D) qPCR measurements of HBV genome copy numbers in supernatants of HepG2.215 cells (control and SENP3 K.D.). HBV 1.3-mer WT replicon plasmid was used as internal control. The data were mean±SD (n = 3) and the statistical significance was assessed by Students’ unpaired t-test. (E) Left: RT-qPCR confirmation of SENP3 knockdown in HepG2-NTCP cells. Right: qPCR measurements of HBV genome copy numbers in supernatants of HepG2-NTCP cells (control and SENP3 K.D.) on Day 7 after HBV infection. Beta-actin (left) and HBV 1.3-mer WT replicon plasmid (right) were used as internal control. The data were mean±SD (n = 3) and the statistical significance was assessed by Students’ unpaired t-test.