Fig. 5.

Sea1 is involved in the regulation of general autophagy. A, wild-type and indicated deletion strains transformed with a plasmid expressing GFP-ATG8 were subjected to nitrogen starvation as described under “Experimental Procedures” and examined under a fluorescent microscope after 20 h of starvation. Scale bar = 5 μm. B, strains were grown as in A. Samples were taken at the indicated time points and analyzed via Western blotting with anti-GFP or anti-PGK1 antibodies. C, autophagic flux was calculated as a ratio (percentage) of free GFP to total GFP signal (combined free GFP and GFP-ATG8) in the corresponding Western blots from B. D, to estimate autophagic induction, the total GFP signal was normalized to the PGK1 signal. Normalized values at time point 0 were set as 1.