Figure 7 | Scientific Reports

Figure 7

From: PKR activity modulation by phosphomimetic mutations of serine residues located three aminoacids upstream of double-stranded RNA binding motifs

Figure 7

PKR dimerization assay. (a) Principle of the PKR dimerization assay based on split nanoluciferase: PKR constructs C-terminally tagged with nanoluciferase SmBit and LgBit fragments are co-expressed in PKR-KO cells. Upon poly(I:C) transfection, PKR dimerization leads to the assembly of active nanoluciferase. (b) Kinetics of nanoluciferase activity measured after substrate addition, for wt PKR, in mock-transfected cells and in cells transfected with poly(I:C) for 2 h before substrate addition. (c) Graphs show the mean and SEM of nanoluciferase activity induction following poly(I:C) transfection in cells that co-expressed wt or mutant PKR linked to the Sm and Lg Bits of nanoluciferase (homodimerization assay). Luciferase activity was measured 5 min after substrate addition. Significance is shown for multiple one-way ANOVA comparisons with WT PKR and with the negative control 4 K. Graphs were generated with Excel 2011 (Microsoft). Statistical analysis was performed using Prism7 (Graphpad).

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