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. 2016 May 31;7:11664. doi: 10.1038/ncomms11664

Figure 3. INTS10 suppresses HBV replication in an IRF3-dependent manner in HepG2.2.15 cells.

Figure 3

(a,b) Levels of phosphorylated (p-) or total proteins in lysates of HepG2.2.15 cells measured by western blot analyses, with GAPDH or Tubulin indicating protein loading in each lane. The cells were transfected with pLV-EGFP-INTS10 vector (INTS10) or pLV-EGFP control vector (Vector) (a), or with INTS10-specific siRNAs (Si-INTS10#1 and Si-INTS10#2) or non-targeting scrambled siRNA controls (Si-Ctrl) (b). (c) Luciferase activity of ISRE reporter plasmids 48 h after co-transfection into cells with INTS10 or control vectors. RLU, relative luciferase units. (d) The mRNA levels of IFNL1 and IFNL2/3 measured by quantitative real-time PCR (qRT-PCR) normalized to human β-actin gene ACTB in cells transfected with INTS10 or control vectors. (e) Luciferase activity of ISRE reporter plasmids 48 h after co-transfection into cells with INTS10-specific or control siRNAs. (f) The mRNA levels of IFNL1 and IFNL2/3 measured by qRT-PCR normalized to ACTB in cells transfected INTS10-specific or control siRNAs. (g) Luciferase activity of ISRE reporter plasmids 48 h after co-transfection into cells with INTS10 or control vectors and an IRF3-specific siRNA (Si-IRF3#1) or non-targeting scrambled siRNA controls (Si-Ctrl). (h) Levels of INTS10 and IRF3 measured by western blot analyses, with GAPDH indicating protein loading in each lane. (i) The mRNA levels of IFNL1 (left) and IFNL2/3 (right) measured by qRT-PCR normalized to ACTB in cells co-transfected with INTS10 or control vectors and IRF3-specific or control siRNAs. (j,k) Levels of HBV DNAs (j), 3.5 Kb pregenomic RNAs (pgRNAs) and 2.4/2.1 Kb Pre-S/S RNAs (k) in cells co-transfected with INTS10 or control vectors andIRF3-specific or control siRNAs measured by qRT-PCR normalized to ACTB. (l,m) Levels of HBsAg (l) and HBeAg (m) in supernatants of cells co-transfected with INTS10 or control vectors and IRF3-specific or control siRNAs measured by enzyme-linked immunosorbent assays (ELISA). All histograms show mean values from three independent experiments; error bars indicate s.d. P values were determined using two-tailed unpaired t-test. *P<0.05, **P<0.01 and ***P<0.001.