FIG 7.

Ulk1 complex does not nucleate under hyperosmotic stress. (A) Wild-type MEFs and T24 cells were cultured in DMEM (control) or amino acid-free DMEM (−AA) for 120 min or with 0.8 M sucrose for 30 min. They were fixed and double immune labeled for WIPI2 (red) and FIP200 (green). The boxed regions are magnified in the insets. The yellow arrowheads indicate puncta double positive for WIPI2/FIP200. Bars, 10 μm. (B) Wild-type MEFs expressing GFP-Ulk1 (green) and T24 cells were cultured as for panel A. They were fixed and immunolabeled for WIPI2 (red) in MEFs and double immunolabeled for Atg16L (red) and Atg13 (green) in T24 cells. The boxed regions are magnified in the insets. The yellow arrowheads indicate puncta double positive for WIPI2/GFP-Ulk1 in MEFs and Atg16L/Atg13 in T24 cells. Bars, 10 μm.