Skip to main content
. Author manuscript; available in PMC: 2010 Jun 25.
Published in final edited form as: J Biol Chem. 2007 Nov 28;283(4):1962–1973. doi: 10.1074/jbc.M706877200

FIGURE 8. Zymolyase treatment and the fps1 mutation cause reductions in wall-associated Ccw12p.

FIGURE 8

Anti-HA Western analysis of Ccw12 protein levels in strains carrying the CCW12-HA expression plasmid pSS1564. A, levels of cell wall-associated Ccw12p in various strains were compared by Western analysis following recovery of surface proteins upon (5 mm) DTT treatment and in extracts prepared from the DTT-treated cell pellet. The α-Hog1 Western and Ponceau S staining show even loading of protein extracts in each lane. Equivalent volumes of the DTT supernatants were loaded in the top panel. Wild type (WT), ccw12Δ (JF2278) carrying pSS1564 (CCW12-HA); ccw12Δ, JF2278 carrying vector; fps1Δ, ccw12Δfps1Δ (JF2442) carrying pSS1564 (CCW12-HA). Cells were treated with 1 unit of zymolyase 100T (US Biological) prior to DTT extraction in lane 6. B, strains were grown to log phase, and 10-fold dilutions were spotted on YPD + 0.5 m sorbitol plates and the same type of plate but containing 25 μg/ml HB or 30 μg/ml CFW. Plates were incubated at 30 °C for 48 h prior to scanning.