TABLE 4.
TLR3-mediated interferon signaling after RNA interference of autophagy components
HEK cells expressing wild type TLR3-HA were transfected with the pISRE-LUC reporter and either a scrambled siRNA control duplex or a distinct siRNA duplex directed to MAPL1LC3β, Beclin 1, or Atg5 and then stimulated with 25 μg/ml poly(I:C) for 24 h. The amount of luciferase produced was measured in the whole cell extracts and expressed as the mean ± S.E. relative to the luciferase activity in unstimulated WT cells. Statistical comparisons of the measurements are listed in the legend to Figure 6.
| Gene target | siRNA duplex | Relative luciferase activity |
|
|---|---|---|---|
| −Poly I:C | +Poly I:C | ||
| MAP1LC3β | |||
| NTa | 1.0 ± 0.1 | 26.7 ± 2.6 | |
| Scrambled siRNA-A | 1.8 ± 0.2 | 31.2 ± 5.2 | |
| WT MAP1LC3β siRNA-A | 0.5 ± 0.1 | 4.8 ± 0.8 | |
| Mutant MAP1LC3β siRNA-A | 0.8 ± 0.1 | 15.0 ± 1.8 | |
| WT MAP1LC3β siRNA-B | 1.5 ± 0.5 | 2.7 ± 0.9 | |
| WT MAP1LC3β siRNA-C | 0.8 ± 0.2 | 1.9 ± 0.6 | |
| Beclin 1 | |||
| NT | 1.0 ± 0.1 | 11.9 ± 1.2 | |
| Scrambled siRNA-A | 1.0 ± 0.1 | 10.1 ± 0.9 | |
| Beclin 1 siRNA-A | 0.3 ± 0.1 | 2.8 ± 0.1 | |
| Beclin 1 siRNA-B | 0.5 ± 0.1 | 2.5 ± 0.2 | |
| Beclin 1 siRNA-C | 0.5 ± 0.1 | 4.7 ± 0.7 | |
| Mutant Beclin 1 siRNA-C | 2.1 ± 0.5 | 9.8 ± 1.9 | |
| Atg5 | |||
| NT | 1.0 ± 0.1 | 19.7 ± 1.6 | |
| Scrambled siRNA-A | 0.9 ± 0.1 | 11.4 ± 1.8 | |
| Atg5 siRNA-A | 0.8 ± 0.1 | 4.2 ± 0.6 | |
| Mutant Atg5 siRNA-A | 0.8 ± 0.1 | 11.1 ± 1.1 | |
| Atg5 siRNA-B | 1.4 ± 0.2 | 10.7 ± 1.8 | |
| Mutant Atg5 siRNA-B | 1.7 ± 0.3 | 20.8 ± 3.2 | |
a NT, non-treated with siRNA WT TLR3 cells were transfected with pISRE-LUC reporter only, whereas treated cells were subjected to two rounds of transfection with a specific siRNA duplex before co-transfection of the siRNA with the luciferase reporter.