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. 2015 Mar 17;6(2):e02486-14. doi: 10.1128/mBio.02486-14

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Copyright © 2015 Reineke et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-ShareAlike 3.0 Unported license, which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original author and source are credited.

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FIG 2 — G3BP1 and Caprin1 directly interact with PKR. (A) Purified inactive PKR (PKR) or active PKR (P-PKR) were incubated in the presence of MBP-tagged MS2 or MBP-tagged G3BP1 deletion mutants, as indicated, and precipitated with amylose resin. Precipitated material was examined with Western blotting for total PKR. (B) MBP precipitations were performed as described for panel A with either MBP-tagged MS2 or G3BP1. Reactions were performed in the presence of ATP with or without poly(I:C) to induce a conformational change in PKR protein. (C) Purified Caprin1 and either inactive PKR (PKR) or active PKR (P-PKR) were incubated together. IPs were performed with protein A-Sepharose and either nonspecific IgG or Caprin1 antibodies, as indicated. Precipitates were analyzed by Western blotting for Caprin1 or total PKR. Relative amounts of PKR and P-PKR inputs are shown with different Western blot exposures (separated by a black line).