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. 2015 Aug 24;10(8):e0136708. doi: 10.1371/journal.pone.0136708

Fig 3. DIV crude supernatant induces rapid IL-1β secretion independent of viral replication.

Fig 3

(A-D) Time course of mobilized monocytes inoculated with mock supernatant or DIV crude supernatant with or without mAb 5G22. Cells were washed at 1 hpi and resuspended in fresh medium. Samples were collected at 2, 8, 16, and 24 hpi. (A) Secreted IL-1β. (B) Intracellular DENV E-protein expression. (C) Infectious virus present in the supernatant, as measured by immunoassay on Vero cells. (D) Relative expression of DENV genome copies in mobilized monocytes, measured by real-time PCR. Values are normalized to 2 hpi samples in the absence of mAb 5G22. For A-D, data are pooled from two independent experiments. (E) Secreted IL-1β by mobilized monocytes at 4 hpi. (F) Secreted IL-1β by fresh, non-mobilized monocytes at 4 hpi. (G) Intracellular DENV E-protein expression at 24 hpi with live DIV crude supernatant or DIV crude supernatant inactivated with formalin or UV exposure, all in the presence of mAb 5G22. (H) Secreted IL-1β at 24 hpi using supernatants from 3G. Tests used: Two-Way ANOVA with Dunnett’s post-test (A–D), One-Way ANOVA with Dunnett’s post-test (G, H), Two-Way ANOVA with Tukey’s post-test (E, F). For A and B, gray (A only) and blue asterisks compare DENV and DENV + 5G22, respectively, to mock within each time point. For C and D, blue asterisks compare DENV + 5G22 to the 2 hpi time point.