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. 2010 Apr 12;207(4):793–806. doi: 10.1084/jem.20092545

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© 2010 Avirutnan et al.

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Figure 2. — DENV NS1 inhibits C4 to prevent CP dependent hemolysis. (A and B) Human C4 was incubated in the presence or absence of DENV NS1 and then added to sensitized EA bearing C1 (EAC1). After three washes, the EAC14b cells were incubated with purified C2, washed, and incubated with GPS treated with EDTA (A) or purified human components (B; C3-C9) at 37°C for 30 min. After centrifugation, the supernatant was analyzed by spectrophotometry at 414 nm. (C) Single-step hemolysis assay. EA were incubated with an 80-fold dilution of C4-deficient GPS that was supplemented with human C4, with or without preincubation with DENV NS1. The data represent the mean ± SD for three independent experiments. Asterisks denote hemolysis conditions that are statistically different from untreated cells (*, P < 0.05; **, P < 0.005; ***, P < 0.0005).