FIG. 2.
Targeting of GFP reporters by full-length forms of Ecm331p and Hwp1p. C. albicans cells expressing the Hwp1/FlagGFP reporter fused to all of Ecm331p or Hwp1p except for the N-terminal signal peptides (Hwp1/FlagGFP/Ecm331FL or Hwp1/FlagGFP/Hwp1FL, respectively) were fluorescent at the cell surface (A), but only the Hwp1/FlagGFP/Hwp1FL reporter was recognized on the surfaces of intact C. albicans cells by antibodies to the Flag epitope (B). (C) The cells expressing each of these proteins were fractionated into lysate (LYS), noncovalently linked CW (SDS), and glucanase-released CW fractions. When Western blots of fractionated cells were probed with antibodies to the Flag epitope, abundant amounts of the Hwp1/FlagGFP/Hwp1FL fusion targeted to the CW, whereas the Hwp1/FlagGFP/Ecm331FL fusion did not. (D) The Hwp1/FlagGFP/Ecm331FL reporter was present in the detergent fraction of Triton X-114 extracts of non-PI-PLC-treated whole-cell lysates, but not in PI-PLC-treated lysates. The arrows indicate the major bands of the GFP fusion proteins, and molecular sizes in kDa are shown at the left.