Extended Data Fig. 7. The catalytically deficient mutant Ded1^DQAD forms constitutive stress granules, without being compromised for RNA or ATP binding.

(A) Ded1^DQAD-mCherry, but not wild-type Ded1-mCherry, forms SGs (marked with arrows) in unstressed cells. Representative images of >3 independent experiments. (B) Fluorescence polarization analysis to measure binding of MANT-ATP to either wild-type Ded1 or Ded1^DQAD. n = 3 technical replicates, mean and SD; nonlinear fit (on site binding curve) calculated using Prism (Graphpad). (C) Fluorescence polarization analysis to measure binding of a fluorescein-UTP labeled, 100bp-long RNA to wild-type either wild-type Ded1 or Ded1^DQAD. n = 3 technical replicates, mean and SD; nonlinear fit (on site binding curve) calculated using Prism (Graphpad).