Figure 3. Dicer and its co‐factors do not impact on the ability of LGP2 to bind viral RNA and induce MDA5‐dependent type I IFN signalling.

1. HEK293 cells stably expressing FLAG‐LGP2 were treated with siRNAs targeting MDA5, TRBP, PACT or Dicer or non‐targeting control siRNAs. Knockdown efficiency was determined by SDS–PAGE and immunoblotting using the indicated antibodies (left panel). β‐Actin serves as loading control. Three days post‐transfection, the IFN response to the indicated doses of total RNA extracted from EMCV‐infected HeLa cells was tested in an IFN‐β promotor luciferase‐based reporter assay (right panel). Mean values and SEM of three independent experiments are shown. Statistical analysis was performed using one‐way ANOVA with Sidak's multiple comparisons test as post‐test for pairwise comparisons using untransfected cells as control for each dose. Significant differences were observed for siMDA5 only (*P < 0.05).
2. HeLa cells were treated with non‐targeting control siRNAs or siRNAs targeting Dicer, transfected with FLAG‐LGP2, and 48 h later infected with EMCV (MOI 1). Twelve hours post‐infection, cells were lysed and LGP2 was retrieved by FLAG immunoprecipitation. The associated RNA was extracted from the immunoprecipitates and tested in the IFN‐β promotor luciferase reporter assay in HEK293 cells at two doses (1× and 5×).

Source data are available online for this figure.