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. 2022 Dec 14;146(7):2897–2912. doi: 10.1093/brain/awac479

Figure 1.

Figure 1

Non-stop mRNA protein products are degraded by the RQC in a Listerin-dependent manner in mammalian cells. (A) Schematic of stalling reporters and EGFP constructs. (B) GFP versus RFP expression profile of stalling reporter with poly-A sequences compared to a no insert control (left). Ratio of median fluorescence intensity of RFP to GFP shows that poly-A sequences induce stalling independent of listerin. Data are shown as mean ± standard error of the mean (SEM) with all data points (two-way ANOVA, n = 3, reporter P < 0.0001, LTN1 KO P = 0.9771, interaction P = 0.8548) (C) Confocal images of HEK 293T and LTN1 KO cells expressing EGFP-NS reporter show accumulation and reduced clearance of NS protein in LTN1 KO cells over time (scale bar = 50 µm). (D) Listerin overexpression in LTN1 KO cells significantly reduces levels of NS reporter protein in cells quantified by flow cytometry. Percentage is shown as mean ± SEM (mixed effects model, n = 3, Ltn1:HEK 293T P = 0.1246, Ltn1:LTN1 KO #1 P = 0.0001, Ltn1:LTN1 KO #2 P < 0.0001).