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Index of Online Supplemental Material for
J. Cell Biol. 10.1083/jcb.200803137
Axe et al.
• Figure S1 Response of three PI(3)P-binding proteins to starvation.
• Figure S2 Overexpression of WT DFCP1 at high levels inhibits autophagosome formation, whereas down-regulation of DFCP1 by RNAi does not inhibit autophagosome formation.
• Figure S3 Colocalization of GFP-DFCP1 with autophagy-related proteins and the ER.
• Figure S4 Localization of DFCP1 by immuno-EM.
• Figure S5 Three independent assays for autophagy in the cell lines used throughout this work.
• Video 1 Double imaging of GFP-FYVE-TM and dsRed-ER during starvation showing formation of GFP-FYVE-TM punctate structures.
• Video 2 Single imaging of GFP-DFCP1 during starvation showing formation of DFCP1 punctate structures.
• Video 3 Triple imaging of GFP-DFCP1 (green), mRFP-MAP-LC3 (red), and CFP-ER (blue) during starvation showing formation of an autophagosomes from an omegasome (arrow).
• Video 4 Double imaging of GFP-DFCP1 (green) and dsRed-ER (red) during starvation (using TIRF) showing omegasome formation and collapse in close association with the ER.
• Video 5 Video showing a 3D reconstruction of GFP-DFCP1 (green) and mRFP-MAP-LC3 (red) at different planes of rotation.
• Video 6 Double imaging of GFP-DFCP1 (green) and mRFP-MAP-LC3 (red) during starvation showing several examples of the last step in autophagosome formation that involves omegasome exit.
• Video 7 Triple imaging of GFP-DFCP1 (green), mRFP-Vps34 (red), and CFP-ER (blue) during starvation showing formation of an omegasome in tight association with a Vps34 particle.
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