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. 2022 Mar 30;5(7):e202201423. doi: 10.26508/lsa.202201423

Figure 6. Nuclear pore complex (NPC) exclusion requires Lem2 and centromere tethering.

Figure 6.

(A) SPA-SIM of Lem2-GFP (yellow) and Ppc89-mCh (magenta). N, number of averaged images. Lem2-GFP and Ppc89-mCh intensity profiles are shown below. Overlay of SPA-SIM datasets for Lem2-GFP (magenta) and Nsp1-GFP (yellow). Bars, 0.5 μm. (B) SPA-SIM Nsp1-GFP (yellow) and Ppc89-mCh (magenta) in wild-type, lem2∆, ima1∆, and man1∆ backgrounds. Bar, 0.5 μm. Plot of NPC exclusion zone dimensions (full width half maximum, ±95% CI). Nsp1-GFP intensity in spindle pole body proximal and distal regions was plotted and compared using a Wilcoxon rank-sum test. Black dots represent the mean normalized intensity value, and error bars show SD. (C) Schematic of Lem2, including rescue constructs. (D) SPA-SIM images of Nsp1-GFP (yellow) in lem2∆ cells with lem2FL, lem2∆C, or lem2∆N constructs turned on or off. Bar, 0.5 μm. Plot of NPC exclusion zone dimensions (full width half maximum, ± 95% CI). (B) For comparison, wild-type and lem2∆ dimensions from (B) are also shown. (E) Schematic of centromere tethering at the spindle pole body in wild-type and csi1∆ cells. SPA-SIM of Nsp1-GFP (yellow) and Ppc89-mCh (magenta) in csi1∆ cells. N, number of averaged images. Bar, 0.5 μm.