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. Author manuscript; available in PMC: 2009 Sep 30.
Published in final edited form as: J Biol Chem. 2001 Oct 23;277(1):763–773. doi: 10.1074/jbc.M109134200

Fig. 8. The perivacuolar compartment is a physiological intermediate in the Cvt and autophagy pathways.

Fig. 8

The apg9Δ pep4Δ strain was co-transformed with a plasmid expressing a temperature conditional allele of Apg9 (pAPG9ts(414)) and the copper-inducible GFP-Aut7 fusion protein (pCuGFPAUT7(416)). The transformed cells were grown at nonpermissive temperature (37 °C) to midlog stage, induced with 10 μm CuSO4 for 2 h, and labeled with FM 4–64 at 37 °C. The cultures were then incubated in SD-N medium for 1 h at 37 °C and viewed directly or shifted to permissive temperature (25 °C) for 30 and 120 min prior to fluorescence microscopy analysis as described in Fig. 1 and under “Experimental Procedures.” GFP-Aut7 fluorescence that accumulates at the PVC under nonpermissive conditions (0 min) can be seen to migrate to punctate structures (30 min) that ultimately end up in the vacuole lumen (120 min) at permissive temperature.