Figure 7.

RcR, RcD, Rc2′F and Dc2′F NANPs stimulate RIG-I dependent responses. (A) HEK-Lucia RIG-I reporter cells were transfected with 5 nM NANPs for 24 h. RIG-I activity was evaluated using Quanti-Luc to monitor luciferase activity. (B) Microglia were treated with scrambled siRNA or siRNA targeting RIG-I (αRIG-I) at a final concentration of 5 nM for 24 h. Cells were placed in fresh media for 24 h prior to transfection with 5 nM NANPs. Cell supernatants and lysates were collected 24 h post transfection. Cell lysates were evaluated for RIG-I protein expression via immunoblot analysis. Cell supernatants were evaluated for IL-6 and IFN-β by specific capture ELISAs. Data are represented as the mean ± (SEM) for a minimum of three independent experimental replicates. Asterisks indicate statistical significance compared to L2K (A) or compared to the corresponding negative control abbreviated as C. (Student's t-test or two-way ANOVA, P-value < 0.05, DcR P-value = 0.0507).