Extended Data Fig. 7: Addition of an unstructured GS-peptide tag accelerates the rate of nuclear shuttling of cargos of large mass and extremely high mechanical stability.
From: Structural anisotropy results in mechano-directional transport of proteins across nuclear pores
(A) Left: Representative confocal images of U2OS cells 30 min in the recovery phase. Scale bar 10 µm. Right: Average time courses of the relative nucleus-to-cytoplasm localization of Spy0128-(WT)2, (GS)8-Spy0128-(WT)2 and (GS)25-Spy0128-(WT)2. The (GS)-tag accelerates the nuclear import kinetics of large protein cargos with extreme mechanical stability. (B) Protein import rate constants (mean ± SEM) calculated from the accumulation curves. Spy0128-(WT)2 (kI = 0.56 ± 0.03 ks−1, n = 104); (GS)8-Spy0128-(WT)2 (kI = 0.83 ± 0.08 ks−1, n = 43); (GS)25-Spy0128-(WT)2 (kI = 0.65 ± 0.04 ks−1, n = 92). Significance levels for two-tailed Mann-Whitney non-parametric test NS P > 0.05, *P ≤ 0.05. Spy0128-(WT)2 vs. (GS)8-Spy0128-(WT)2, P = 0.01; (GS)8-Spy0128-(WT)2vs. (GS)50-Spy0128-(WT)2, P = 0.07; Spy0128-(WT)2 vs. (GS)50-Spy0128-(WT)2, P = 0.21. All points and bar plots indicate mean ± SEM.
