Figure 5.

cGAS preferentially interacts with collided ribosomes

(A) Strategy to generate collided ribosomes using an in vitro translation reaction in rabbit reticulocyte lysate (Juszkiewicz et al., 2018).

(B) cGAS was incubated with a mixture of collided and non-collided ribosomes at different ratios, separated by 10%–50% sucrose gradient fractionation, and then analyzed by western blotting. Collided ribosomes (fractions 8, 9, and 10) are indicated by dashed lines.

(C) Western blot analysis of MRC5VA cytosol fractionated by sucrose gradient sedimentation, with or without prior incubation with micrococcal nuclease (MNase) to digest polysomes (poly) to monosomes (mono).