Skip to main content
. 2010 Aug 18;6(6):764–776. doi: 10.4161/auto.6.6.12709

Figure 6.

Figure 6

ULK1 puncta tightly associate with the ER membrane. (A and B) NIH3T3 cells stably expressing GFP-ULK1 (A) or Vps34-GFP (B) together with HA-Atg14 were cultured in starvation medium with 0.2 µM wortmannin for 1 hour. Cells were then fixed, permeabilized, and subjected to immunofluorescence microscopy using anti-HA and anti-GFP (for GFP-ULK1) antibodies. signal color is indicated by color of typeface. St. M., starvation medium; WM, wortmannin. Scale bars, 10 µm (white) and 1 µm (yellow). (C) MEFs stably expressing GFP-ER (containing transmembrane region of rat cytochrome b5) and HA-ULK1 were cultured in starvation medium, with or without 0.2 µM wortmannin for 1 hour, and then subjected to immunofluorescence microscopy using anti-GFP and anti-HA antibodies. (D and E) selected frames from time-lapse movies of MEFs stably expressing GFP-ULK1 and mRFP-ER in starvation medium, with (E) or without (D) 0.2 µM wortmannin are shown. The cells were imaged starting from 30 minutes (D) or 90 minutes (E) after the starvation treatment commenced. Localization of GFP-ULK1 is indicated by arrowheads. See Video 1 and 2 for whole images. Signal color is indicated by color of typeface. St. M., starvation medium; WM, wortmannin. Scale bars, 10 µm (white) and 1 µm (yellow).