Supplemental Material

This article contains the following supporting material:

• Supplemental Figure 1 - GFP-Ape1 is targeted to the PAS and delivered to the vacuoles in wild type cells. Wild type cells (SEY6210) harboring the plasmid to express GFP-Ape1 were grown to mid-log phase, labeled with FM 4-64, and immediately examined (SMD), or after further cultured in nitrogen starvation medium for 2 to 3 h (SD-N), by fluorescence microscopy. The GFP signals were shown in green color and FM 4-64 signals in red.
• Supplemental Figure 2 - Atg11 is co-isolated with Atg9 in atg2Δ, atg17Δ, atg18Δ, or atg20Δ cells. Protein extracts from the indicated cells expressing ProtA-Atg9 and myc-tagged Atg11 were incubated with human IgG-coated magnetic beads. Co-isolated proteins were subjected to SDS-PAGE followed by immunoblotting with anti-myc antibody and PAP reagents for detecting protein A-tagged Atg9.
• Supplemental Figure 3 - Starvation treatment stimulates Atg9 restriction to puncta away from mitochondria in atg1Δ and atg1Δ ape1Δ cells. Mitochondria of the indicated strains were labeled with GFP (mitoGFP). Their colocalization with RFP-Atg9 were examined in cells grown to mid-log phase (SMD), or after further cultured in nitrogen starvation medium for 2 to 3 h (SD-N), by fluorescent microscopy.
• Supplemental Figure 4 - Atg11 is targeted to the perivacuolar PAS normally in the absence Atg9. Cells of the atg1Δ atg9Δ strain co-expressing GFP-Atg11 and the indicated Atg9 variants or harboring an empty vector were grown to mid-log phase, labeled with FM 4-64, and examined immediately (SMD) , or after further cultured in nitrogen starvation medium for 2 to 3 h (SD-N), by fluorescence microscopy.