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. 2022 Apr 14;33(5):ar35. doi: 10.1091/mbc.E21-12-0616

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© 2022 Vitale, Khan, et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.

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FIGURE 5: — Nsp1 localizes at the base of the P_Gal1-brl1^F391E-induced petals. (A–E) Cells expressing (A) yeGFP, (B) BRL1-yeGFP, and (C) brl1^F391E-yeGFP for 3 h at 30°C were analyzed for the localization of yeGFP or the yeGFP-tagged Brl1 and Nsp1 by immuno-EM. (A) Magenta arrowheads point toward Nsp1 at NPCs. (B) Green arrowheads point toward Brl1-yeGFP at membrane sheets in the nucleus, and magenta arrowheads point toward Nsp1 at NPCs. (C) Green arrowheads highlight Brl1^F391E in NE attached petals (i and ii). Nsp1 is found beside NPCs on the base of the NE-associated petals (iii and iv; magenta arrowheads). Enlargements from cells i–iv show NE-associated petals with Brl1 (green) or Nsp1 signal (magenta) at the base of the petals. (A, B) Scale bars: 200 nm. (C) Scale bar: 500; enlargements: 200 nm. (D, E) Quantification of gold particles reflecting the localization of brl1^F391E-yeGFP (D) or Nsp1 (E) from 9 and 11 cells, respectively. Abbreviation: N, nucleus.