Figure 4

Intracellular s-RNYs activate TLR7 to promote apoptosis and inflammation in monocytes/macrophages. Cell death data by flow cytometry from human THP1 cells (a – left panels) or mouse BMDMs (b – left panels) transfected with a cocktail of siRNAs against the terminal loop of all RNYs (siRNYs) to induce s-RNY maturation or control (siCTL). Cells were incubated with 50 mM of chloroquine for 48 h (left panels) or with the TLR7/9 specific inhibitor IRS954 at 10 μg/ml of concentration for 48 h (middle panels). Percentage of apoptotic cells was determined by staining with annexin V-FITC. Data are presented as mean and S.D. (n=3). The same experimental conditions were analyzed by immunoblot of IκBα, actin, total caspase 3 (casp3) and its cleaved form (c-casp3) in THP1 cells (a – right panel) or BMDMs (b – right panels). Student’s t-test: *P<0.05; ***P<0.001