Immunofluorescence (IF) analysis of RIG-I, MAVS, and dsRNA (red) with G3BP1 (green) in U2OS cells. SeeFigure S1A for antibody validation. Cells were transfected with 162 bp dsRNA containing 5ppp (500 ng/mL) for 6 h before imaging. Raw images for RIG-I and MAVS without contrast adjustment (raw) were also shown. For dsRNA imaging, 162 bp dsRNA 3-labeled with Cy5 was introduced into cells by lipofectamine transfection or electroporation. Unless mentioned otherwise, unlabeled dsRNA and lipofectamine transfection was used throughout the manuscript. Cell nuclei were stained with Hoechst 3342. Bottom right: SG colocalization was measured by Pearson colocalization coefficient (PCC) between G3BP1 foci and indicated molecules from 10 fields of view.
