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. 2017 Mar 29;91(8):e01824-16. doi: 10.1128/JVI.01824-16

FIG 1.

FIG 1

MMP-9 is upregulated in PBMCs of CHB patients and activated by HBV in PBMCs and macrophages in vitro. (A) MMP-9 mRNA levels in PBMCs of CHB patients (n = 69) and healthy individuals (n = 40) were measured by qPCR. Points represent MMP-9 mRNA levels of each sample. (B to G) PBMCs (1 × 106) obtained from healthy individuals were incubated with supernatants isolated from HepG2 cultures (without HBV) or HepG2.2.15 cultures (containing HBV) at an MOI of 0.5 for different times (B to D) or for 24 h at different MOIs (E to G). The MMP-9 mRNA level was measured by qPCR and normalized to the GAPDH mRNA level. (B and E) Results are standardized to a value of 1 for HepG2 supernatant-treated cells. (C and F) MMP-9 proteinase activity in the supernatants was determined by gelatin zymography assays. (D and G) The MMP-9 protein level in WCLs was determined by Western blotting. (H to M) Macrophages (1 × 106) differentiated from THP-1 cells were incubated with supernatants isolated from HepG2 cultures or HepG2.2.15 cultures at an MOI of 0.5 for different times (H to J) or at different MOIs for 48 h (K to M). The MMP-9 mRNA level was measured by qPCR and normalized to the GAPDH mRNA level. (H and K) Results are standardized to a value of 1 for HepG2 supernatant-treated cells. (I and L) MMP-9 proteinase activity in the supernatants was determined by gelatin zymography assays. (J and M) The MMP-9 protein level in WCLs was determined by Western blotting. (N) PBMCs (1 × 106) isolated from healthy individuals were incubated with supernatants of HepG2 cultures, supernatants of HepG2.2.15 cultures, anti-HBsAg antibody-pretreated supernatants of HepG2.2.15 cultures, or UV-inactivated HepG2.2.15 supernatants. The MMP-9 mRNA level was measured by qPCR. (O) Macrophages (1 × 106) differentiated from THP-1 cells were incubated with supernatants of HepG2 cultures, supernatants of HepG2.2.15 cultures, anti-HBsAg antibody-pretreated supernatants of HepG2.2.15 cultures, or UV-inactivated HepG2.2.15 supernatants. The MMP-9 mRNA level was measured by qPCR. (P) PBMCs (1 × 106) isolated from healthy individuals were incubated with supernatants of HepG2 cultures, supernatants of cultures of HepG2 cells transfected with pHBV1.3, supernatants of cultures of HepG2 cells transfected with pCMV-HBS, or supernatants of HepG2.2.15 cultures. The MMP-9 mRNA level was measured by qPCR. Graphs show means ± standard deviations (n = 3). *, P < 0.05; **, P < 0.01; ***, P < 0.001.