Figure 2 .

Knocking out PKR and PERK kinases decreases DM2 RAN protein levels. CAGG (A) and CCTG (B) repeat expansion constructs used to co-transfect WT and KO HEK293T cell lines with a GFP expressing control plasmid. These constructs have two upstream stop codons and a C-terminal epitope tag in each reading frame. Protein blots showing QAGR (A) or LPAC (B), PKR, p-eIF2α, total eIF2α, GFP and GAPDH levels in WT versus PKR KO (lower left) or WT versus PERK KO cells (lower middle). Quantification using ImageJ software shows normalized QAGR/GAPDH (A) or LPAC/GAPDH ratios (B) (lower right). Error bars show SD. n = 3. Statistical analyses were performed using the two-tailed t-test, ^*P < 0.05, ^**P < 0.01, ^****P < 0.0001.