Figure 4. Loss of SIDT2 impairs endosomal escape of internalised poly(I:C).

BMDCs from Sidt2^+/+ and Sidt2^−/− mice were treated with (A) poly(I:C)-rhodamine and (B) ³²P-labeled 500bp dsRNA for 60 min at either 4°C or 37°C, and internalisation assessed via flow cytometry or radioactivity measurement respectively. Results are representative of at least 2 independent experiments. For panel B, all treatments and measurements were made in triplicate, and data are plotted as mean ± SEM. (C–D) BMDCs from Sidt2^+/+ and Sidt2^−/− mice were treated with poly(I:C) for either 10 or 60 min, stained with J2 anti-dsRNA antibody (red) and DAPI (blue), and imaged by confocal microscopy. The proportion of each cell occupied by punctate dsRNA staining was quantified. (E–F) Sidt2^+/+ and Sidt2^−/− BMDCs were treated with poly(I:C) in association with the cationic polymer DEAE-dextran for 60 min. For panels C and E, images are representative of at least three independent experiments. For panels D and F, data are plotted as mean ± SEM and between 30–150 cells were assessed per time point. ^* P < 0.05. Scale bar = 10 μm. See also Figure S5.