Figure 8. Association of actin with PPP1R15A promotes eIF2α phosphatase activity in vitro.
(A) Silver-stained SDS-PAGE or GFP-affinity purified PPP1R15A-GFP (hR15-GFP) and associated proteins. Asterisks signify identity confirmed by mass spectrometry. Purified complex incubated with phosphorylated recombinant eIF2α N-terminal lobe (eIF2α) for incubated time. Note size shift corresponds to dephosphorylation. (B) Quantification of ‘A’ using ImageJ software. Mean ± SEM. p value calculated by two-way ANOVA, n = 3. (C) Immunoblot for PPP1R15A and eIF2α of HEK293T cells expressing hPPP1R15A-GFP (hR15A-GFP). Treated with cycloheximide 50 µM for indicated times. (D) Quantification of ‘C’. Mean ± SEM, n = 3.