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. 2004 Aug;15(8):3553–3566. doi: 10.1091/mbc.E04-02-0147

Figure 7.

Figure 7.

Lipidation of Atg8 is reduced in atg21Δ cells. (A) Atg8 lipidation in rich medium versus nitrogen starvation medium. Cells were grown in YPD to early mid-log phase and then starved for 3 h in SD-N. Atg8-PE was separated from Atg8 by 12% SDS page gels containing 6% urea (Kirisako et al., 2000). All strains are of BY4742 background. (B) Deletion of ATG21 antagonizes the accumulation of Atg8-PE in autophagy mutants. WT (SEY6210), atg21Δ (PSY7), atg18Δ (JGY3), atg18Δ atg21Δ (PSY167), atg9Δ (JKY007), and atg9Δ atg21Δ (PSY191) cells were starved for 4 h in SD-N, and Atg8-PE was analyzed by Western blot. (C) Deletion of ATG21 antagonizes the accumulation of GFP-Atg8 at the PAS in autophagy mutants. Strains used in B containing a plasmid expressing GFP-Atg8 from the CUP1 promoter were grown to mid-log phase in SMD and then starved for 4 h in SD-N. (D) atg21Δ cells have a reduced rate of Atg8 lipidation. Cells of BY4742 background were labeled and Atg8 immunoprecipitated as described in MATERIALS AND METHODS and analyzed by 12% SDS-urea gels.