Figure 7. Loss of Dicer triggers the interferon response.

a) Western blot analysis of cell extracts isolated from normal and Dicer knockdown HEK293 cells, cultured for 1 or 2 weeks, using antibodies specific to Dicer, TLR3, PKR1 and β-actin. Anti-TLR3 antibody detects two bands and according to size markers, the upper band is likely to correspond to modified protein. Quantitation is shown in Supplementary Figure 7. See Supplementary Fig. 9 for uncropped blot images.

b) Western blot analysis of cell extracts isolated from normal and Dicer knockdown HEK293 cells, cultured for 10 days using antibodies specific to IFN-β, OAS1 and tubulin. See Supplementary Fig. 9 for uncropped blot images.

c) Western blot analysis as in (b) using antibodies specific to ADAR1 and tubulin. See Supplementary Fig. 9 for uncropped blot images.

d) Western blot analysis of cell extracts isolated from normal and siRNA-directed Dicer knockdown HEK293 cells using antibodies specific to OAS1, TLR3 and tubulin. See Supplementary Fig. 9 for uncropped blot images.

e) Western blot analysis of cell extracts isolated from normal and Drosha knockdown HEK293 cells using antibodies specific to Drosha, TLR3 and tubulin. See Supplementary Fig. 9 for uncropped blot images.

f) Flow cytometry based quantitation of the % of cell population undergoing apoptosis in normal and Dicer, Drosha or PKR knockdown cells. % of cell population values are based on average values ± s.e.m. from three independent biological experiments. All experiments in the figure were independently repeated 3 times.