Figure 1.
CHMP7 Is an ER-Localized Protein that Is Enriched at the NE during Mitotic Exit
(A) Schematic depicting constructs used in this study.
(B and C) CAL-51 cells edited to express mNG-CHMP7 were resolved and examined by western blotting with anti-CHMP7 and anti-Vinculin (B) or imaged live (C). In this and all other figures, endogenous CHMP7 is marked by an arrowhead (∗, non-specific band).
(D–F) HeLa cells stably expressing GFP-CHMP7 and GFP-CHMP7NT were imaged live (D), lysed, resolved, and examined by western blotting with anti-GFP, anti-CHMP7, or anti-GAPDH antisera (E) or fixed and stained with anti-Calnexin and DAPI (F). Images in (D) are representative of all cells imaged and 22/22 (GFP-CHMP7) and 21/21 (GFP-CHMP7NT) captured movies. Co-localization of GFP-CHMP7 and GFP-CHMP7NT with Calnexin was observed in 7/7 and 13/13 scored cells, respectively.
(G) Post-nuclear supernatants from Cos7 cells were fractionated through a continuous iodixanol gradient and analyzed by western blotting with the indicated antisera.
(H) HeLa cells stably expressing GFP-CHMP7 δNT were imaged live. Images are representative of all cells imaged and 5/5 captured movies. Time interval is presented in seconds post-cortical ingression.
In all micrographs, the scale bar represents 10 μm. See also Figure S1.