Fig. 5

Screening of HOPS engineered strains with disrupted proteases. Detection of secreted HyHEL-Fab (a) or CES (b) by ELISA and Western blot. Prior to loading, the protein amounts were normalized to the wet cell weight of each strain. The relative fold changes of extracellular HyHEL-Fab yields (a) were calculated from 4 independent screenings with 3–6 clones of ∆ypt7∆pep4, ∆ypt7∆vps70, ∆vam6∆pep4, ∆vam6∆vps70 and Fab#34 in each screening. For CES secretors (b), the relative fold change yields were calculated from one screening with 4–6 clones of ∆ypt7∆pep4, ∆ypt7∆vps70, ∆ypt7∆prb1, ∆vam6∆prb1 and CES#18 (except ∆vam6∆pep4 (2 biological replicates) and ∆vam6∆vps70 (1 clone). Error bars indicate SEM. Statistical significance determined by the Student’s t-test is indicated with an asterisk (p < 0.05*, p < 0.01**, p < 0.001***)