Atg9 vesicles are an important membrane source during early steps of autophagosome formation

Video 7.
In response to starvation, cytoplasmic Atg9 vesicles assembled to the PAS. Video associated with Fig. S5 B. ATG9-2×GFP atg1^D211A atg11Δ VPH1-TagRFPT cells and ATG9-2×GFP atg1^D211A atg11Δ atg17Δ cells were mixed and then treated with rapamycin for 2 h. Images were analyzed by time-lapse microscopy at 30 ms/frame using an inverted microscope (IX71). In atg1^D211A atg11Δ VPH1-TagRFPT cells, Atg9-2×GFP vesicles accumulated at the PAS, and the number of cytoplasmic mobile Atg9 vesicles was decreased. Furthermore, no large clusters were observed in atg1^D211A atg11Δ atg17Δ cells.