Figure 6. Expression of full‐length LGP2 inhibits dsRNA‐mediated RNAi in Ifnar1 ^−/− cells.

1. Ifnar1 ^−/− iLGP2 cells, which also express a destabilised form of GFP (d2GFP), were transfected with Cy5‐labelled long dsRNA corresponding to the first 200 nt of Renilla luciferase (dsRNA‐RL) or GFP (dsRNA‐GFP) in the absence or presence or doxycycline. Forty‐eight hours post‐transfection, cells were harvested and d2GFP expression in live, single, Cy5⁺ cells was analysed by flow cytometry. Histogram plots of two representative clones are shown. Bar graphs display the percentage of GFP median fluorescence intensity of dsRNA‐GFP‐transfected cells relative to dsRNA‐RL‐transfected cells in four independent clones.
2. Unlike full‐length LGP2, LGP2 CTD or CTD K634E expression is unable to suppress dsRNAi in Ifnar1 ^−/− MEFs. Four independent clones of Ifnar1 ^−/− d2GFP MEFs that inducibly express LGP2 CTD or LGP2 K634E were treated with dsRNA‐RL or dsRNA‐GFP as in (A). Histogram plot of one representative clone for each construct is shown. Bar graphs display the percentage of GFP median fluorescence intensity of dsRNA‐GFP‐transfected cells relative to dsRNA‐RL‐transfected cells in four independent clones.

Data information: All histogram plots are representative of three independent experiments. Each histogram represents a sample size of 10,000 cells. Bar graphs depict the median fluorescence values normalised to those in Renilla‐transfected samples. Mean values and SD of three independent experiments are shown. Statistical analysis was performed using two‐way ANOVA with Sidak's multiple comparisons test as post‐test for pairwise comparisons. Significant differences with Sidak's multiple comparisons test are shown (ns, not significant; *P < 0.05; **P < 0.01; ****P < 0.0001).