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. 2022 May 6;221(7):e202111100. doi: 10.1083/jcb.202111100

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© 2022 Zappa et al.

This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/).

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Figure S5. — Suppression of PKR clustering enhances signaling. (A) Image quantification showing that the mutations in PKR’s front-to-front (FTF) kinase interfaces significantly reduce the number of PKR clusters in cells. The data were binned and are shown as the mean and 95% confidence interval bands (n = 2,000 cells). (B) Immunoprecipitation (IP) analysis of the extent of phosphorylation of mRuby-PKR and FTF mutant mRuby-PKR upon 90-min poly I:C treatment. (C) Same as B for FKBP-PKR and FTF mutant FKBP-PKR upon 60 min of dimerizer treatment. (D) Flow cytometry histograms showing the relative fluorescent intensity of cells expressing wild-type mRuby-PKR (blue trace) and FTF mutant mRuby-PKR (red trace). (E) Quantification of nuclear ATF4 signal in immunofluorescence analyses carried out in H4 cells expressing mRuby-PKR or FTF mutant mRuby-PKR treated with poly I:C. (N = 3 experiments, n > 600; **, P < 0.01, unpaired Student’s t test, nonparametric). Source data are available for this figure: SourceData FS5.