Figure 1. Atg39 accumulates at the ONM.... | Rockefeller University Press

Figure 1.

Atg39 accumulates at the ONM.(A) Schematic of localization and topology of split-GFP constructs. The GFP11–mCherry reporter fusion proteins (i, ii, and iii) are constructed as shown in the key, with localization diagrammed. At the bottom are schematics of the Atg39 GFP1–10 fusions and truncations lacking N- or C-termini. The Atg39 TM domain is depicted as a gray oval. Numbers indicate amino acids. (B) Deconvolved fluorescence micrographs of cells expressing indicated Atg39 fusion proteins and mCherry-reporters (see inset). GFP, mCherry, and merged images are shown. Arrows point to NE blebs. Scale bars are 5 μm. (C) Deconvolved fluorescence micrographs of cells expressing GFP1–10-Atg39 and indicated mCherry-reporter (inset). GFP, mCherry, and merged images are shown. Scale bars are 5 μm. Asterisks indicate vacuolar autofluorescence. (D) Normalized line profiles of GFP (green) and mCherry (magenta) fluorescence bisecting cells as indicated by dotted lines in corresponding top and bottom merge panels from C. Position of NE and cER is indicated by dotted lines. (E) As in C but with cells expressing GFP1–10-atg39-(139–398). (F) As in D but with cells from E. (G) As in C but with cells expressing GFP1–10-atg39-ΔL. Asterisks indicate vacuolar autofluorescence. (H) As in D but with cells from G.

Atg39 accumulates at the ONM. (A) Schematic of localization and topology of split-GFP constructs. The GFP¹¹–mCherry reporter fusion proteins (i, ii, and iii) are constructed as shown in the key, with localization diagrammed. At the bottom are schematics of the Atg39 GFP^1–10 fusions and truncations lacking N- or C-termini. The Atg39 TM domain is depicted as a gray oval. Numbers indicate amino acids. (B) Deconvolved fluorescence micrographs of cells expressing indicated Atg39 fusion proteins and mCherry-reporters (see inset). GFP, mCherry, and merged images are shown. Arrows point to NE blebs. Scale bars are 5 μm. (C) Deconvolved fluorescence micrographs of cells expressing GFP^1–10-Atg39 and indicated mCherry-reporter (inset). GFP, mCherry, and merged images are shown. Scale bars are 5 μm. Asterisks indicate vacuolar autofluorescence. (D) Normalized line profiles of GFP (green) and mCherry (magenta) fluorescence bisecting cells as indicated by dotted lines in corresponding top and bottom merge panels from C. Position of NE and cER is indicated by dotted lines. (E) As in C but with cells expressing GFP^1–10-atg39-(139–398). (F) As in D but with cells from E. (G) As in C but with cells expressing GFP^1–10-atg39-ΔL. Asterisks indicate vacuolar autofluorescence. (H) As in D but with cells from G.