Fig. 6.

Flo11p is both shed from the cell wall and covalently attached to it and is expressed and localized similarly in wild-type and vps27Δ strains. Western blotting was performed on fractionated samples from wild-type, vps27Δ, vps25Δ, and rim13Δ strains carrying Flo11-HA^30,1015. A high-molecular-mass Flo11p-HA^30,1015 band (>260 kDa) was observed in wild-type and vps27Δ strains in all fractions, including shed (S), membrane bound/noncovalently cell wall associated (M), and covalently attached to cell wall (C) fractions. The vps25Δ and rim13Δ mutants show the absence of Flo11p-HA^30,1015 in S and C fractions and considerably decreased signals in the M fraction. A small N-terminal fragment (17 kDa) referred to as N-HA was consistently observed in the M fraction.