FIG. 7.

Analysis of HA-tagged Sed1p. The cells containing pRS426-HA::SED1 were cultured in YPAD medium with shaking at 30°C for 30 h. The cell wall fraction was prepared and treated with SDS to remove noncovalently bound proteins. Cell wall proteins were solubilized with RPI or laminarinase and applied to SDS-PAGE (5 to 20% gel). Proteins were transferred to a membrane and probed with anti-HA monoclonal antibody and anti-mouse immunoglobulin G secondary antibody conjugated with alkaline phosphatase. RPI, HA-Sed1p obtained by RPI extraction; LAM, HA-Sed1p obtained by laminarinase extraction.