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. 2022 Feb 26;31(15):2521–2534. doi: 10.1093/hmg/ddac021

Figure 1.

Figure 1

RAN translation is suppressed in vitro in eIF2A KO cell lysates. (A) Schematic of nanoluciferase (NLuc) reporter mRNAs. 3xF = 3x FLAG tag, GA = glycine–alanine, GP = glycine-proline, GR = glycine-arginine, polyG = the glycine reading frame product of the CGG repeat; polyA = the alanine reading frame product of the CGG repeat. (B) Western blot of WT and eIF2A KO lysates probed with an anti-eIF2A antibody to confirm loss of eIF2A expression. GAPDH is used as a loading control. (C) Expression of indicated NLuc reporter mRNAs in eIF2A KO lysates, relative to WT lysates, after controlling for the difference in AUG-NLuc expression between paired lysates, n = 12–42. Graphs represent mean with error bars ± standard deviation. *P < 0.05, **P < 0.01, ****P < 0.0001, non-parametric analysis of variance, Kruskal–Wallis test, with Dunn’s multiple comparison test.