Figure 3.

CSs Are Found at Sites of Autophagosome Formation

(A) HEK293A cells stably expressing GFP-WIPI2b starved for 2 hr in EBSS, fixed and labeled with mouse anti-PCM1 and rabbit anti-GABARAP antibodies. The scale bars represent 5 μm. Arrows indicate triple colocalization; arrowhead indicates GABARAP-PCM1 double colocalization.

(B) HEK293A cells, or MEF cells stably expressing GFP-DFCP1, starved for 2 hr in EBSS, fixed and labeled with the indicated antibodies. The scale bars represent 5 μm. Arrows indicate colocalization or points of contact (GFP-DFCP1) between PCM1 and autophagy markers. Hoechst DNA staining is shown in blue in the merge. In all panels, rabbit anti-PCM1 antibody was used; mouse anti-WIPI2 or LC3B and guinea pig anti-p62 antibodies were used in the remaining panels.

(C) HEK293 cells stably expressing GFP-DFCP1 starved for 2 hr in EBSS followed by immunoisolation with control immunoglobulin G (IgG) or anti-GFP and immunoblot analysis with the indicated antibodies.

See also Figure S2.