FIG 5.
Poly(I·C) induces mitochondrial ROS in polarized 16HBE14o− cells in an NLRX-1- dependent manner. (A) Cells were treated with 1, 5, or 10 μg/ml poly(I·C) and incubated for 6 h at 37°C. Mitochondrial ROS was determined by flow cytometry as described for Fig. 3. (B and C) Polarized monolayers of 16HBE14o− cells were incubated with 0, 10, or 50 μM Mito-Tempo for 1 h both apically and basolaterally and then treated with 5 μg/ml poly(I·C) in the presence of 0, 10, or 50 μM Mito-Tempo. After 6 h, RT was measured, and then the cells were treated with MitoSox Red to determine the levels of mitochondrial ROS. (D and E) Cells were transfected with NT or NLRX-1 siRNA, allowed to polarize for 2 days, and then treated with 5 μg/ml poly(I·C). After 6 h, RT was measured, and then the cells were treated with MitoSox Red to determine the levels of mitochondrial ROS. The data represent means and SD calculated from 3 independent experiments (*, P ≤ 0.05; ANOVA).