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. 2017 Aug 1;7:6975. doi: 10.1038/s41598-017-07308-3

Figure 4.

Figure 4

Antibodies against modified NS1-WD peptide reduce viral replication by complement-dependent cytolysis of DENV-infected cells. (A) HUVECs were infected with DENVs (serotypes 1–4, moi = 10) or mock infection for 48 h. After 48 h of infection, cells were incubated with either PBS, purified pAbs from KLH or modified NS1-WD-peptide immune sera (50 μg/ml) or anti-NS1 mAb 2E8 (50 μg/ml) for 1 h at 4 °C and incubated with or without complement (1:20) for 4 h at 37 °C. The release of lactate dehydrogenase (LDH) was analyzed as described in the Methods. Anti-NS1 Ab 2E8 served as a positive control. (B) HUVECs were infected with DENVs (serotypes 1–4, moi = 10) or mock-infected for 48 h. After 48 h of infection, cells were incubated with either PBS, purified pAbs from KLH or modified NS1-WD peptide-immunized mouse sera (50 μg/ml) or anti-NS1 mAb 2E8 (50 μg/ml) for 1 h at 4 °C and incubated with complement (C’) (1:20) for 4 h at 37 °C. After refilling with fresh medium for another 24 h, the infectious virus in supernatants was titrated by FFA as described in the Methods. All data are presented as the mean ± S.D. from at least three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ns indicates no significance.