Figure EV4. Analysis of doxycycline‐inducible expression of full‐length FLAG‐LGP2, FLAG‐LGP2 CTD, or FLAG‐LGP2 CTD K634E in Ifnar1 ^−/− MEFs.

1. Relative expression (RE) of LGP2 (DHX58) in wild‐type MEFs treated for 24 or 48 h with type I IFN was assessed by qRT–PCR and normalised to β‐actin (ACTB) using the ∆∆C _t method.
2. Verification of doxycycline‐dependent induction of FLAG‐LGP2 expression in Ifnar1 ^−/− MEFs by flow cytometry. Various Ifnar1 ^−/− iLGP2 clones were treated for 72 h with doxycycline (dox) and subsequently fixed, permeabilised and stained with a FLAG‐Cy3 antibody followed by flow cytometry.
3. Verification of doxycycline‐dependent induction of FLAG‐LGP2 CTD and CTD K634E expression in Ifnar1 ^−/− MEFs. The indicated clones were treated for 72 h with dox and subsequently fixed, permeabilised, stained with a FLAG‐Cy3 antibody and analysed by flow cytometry.
4. Immunoblot analysis of four clones of Ifnar1 ^−/− MEFs in which expression of FLAG‐tagged human LGP2 CTD or CTD K634E is induced following 72 h of dox treatment. β‐Actin serves as loading control.
5. Northern blot analysis of dsRNA‐derived siRNAs in two individual clones of Ifnar1 ^−/− iLGP2 CTD and CTD K634E MEFs left untreated or treated with dox for 24 h prior to transfection with dsRNA‐GFP. Twenty‐four hours post‐transfection, cells were harvested and the generation of siRNAs was analysed by Northern blotting using a probe specific for dsRNA‐GFP. The arrow points to dsRNA‐GFP‐derived siRNAs. A miRNA ladder was used as a size marker and endogenous U6 served as loading control.

Source data are available online for this figure.