Figure 5.

Blocking Panx1 or Casp4(11) with sdRNAi in TSP1^−/− LGs decreases LG inflammation. Two to 3 μL solution of Panx1 or Casp4(11) sdRNAi (at a final concentration of 250 nM) was injected into the LGs on the right side of 4-month-old TSP-1^−/− mice. The left LG was injected with a control sdRNAi or vehicle (saline). Mice were injected once a week for the following 5 weeks and euthanized 2 weeks after the last injection. LGs were processed for qRT-PCR or histology. (A) LG treated with specific sdRNAi (to Panx1 or Casp4[11]) showed significant reduction in Panx1 or Casp4(11) mRNA expression compared with LG treated with control sdRNAi. (B) Decrease of Panx1 mRNA level resulted in a significant reduction of expression of both IL1β and Nlrp3, whereas blocking Casp4(11) significantly reduced only IL1β but not Nlrp3 expression. (C) Measurement of the cumulative area of inflammation (summed area of infiltration foci per field, measured in mm²) showed that Panx1 or Casp4(11) sdRNAi treatments of the LG of TSP1^−/− mice significantly decreased the cumulative area of inflammation compared with controls. To quantify statistical difference, a two-tailed Student's t-test was used. Panx1 sdRNAi (P = 1.6 E⁻⁶) and Casp4 sdRNAi (P = 4.0E⁻⁵).