Immunoblotting Protocol for Protein Analysis

Immunoblotting was performed with antibodies against the following proteins: ISG15 (1:1,000; mouse, 703132, clone 1H9L21; human and rat, PA5-88262; Thermo Fisher Scientific), GAPDH (1:1,000; 2188, Cell Signaling Technology), cGAS (1:1,000; 31659, Cell Signaling Technology), STING (1:1,000; 13647, Cell Signaling Technology), RIG-I (1:1,000; 3743, Cell Signaling Technology), MAVS (1:1,000; 4983, Cell Signaling Technology), β-actin (1:10,000; A1978, MilliporeSigma), filamin-C (1:1,000; NBP1-89300, Novus Biologicals), vinculin (1:1,000; 4650, Cell Signaling Technology), p62 (1:1,000; 109012, Abcam), and LC3 (1:1,000; 12741, Cell Signaling Technology). Immunoblotting for the soluble and insoluble fractions of filamin-C was performed in mouse hearts as previously described (82 (link)). Densitometry was performed using ImageJ version 1.39 (NIH).