FIG. 2.

Priming of cells for synergistic expression of cytokines and chemokines occurs with either PGN or poly(I:C). RAW 264.7 cells were treated in tissue culture with either PGN (1 μg/ml), poly(I:C) (10 μg/ml), or both PGN and poly(I:C) either simultaneously or sequentially for 5–10 h. For sequential incubations, cultures were incubated with the first reagent for 5 h, washed with PBS, and then incubated for an additional 5 h with the second reagent. Real-time PCR was performed using GAPDH as a control gene. The average expression of combined treatment with PGN plus poly(I:C) for 5 h was set to 100. Error bars represent mean ± SD. P values for the four treatment groups calculated by ANOVA are indicated separately for each set of culture conditions (n = 3 replicates/condition). Depicted is a representative example from among three repeat experiments.