FIG 3.

SNP+SHAM-induced cell wall changes do not induce Mkc1 or Hog1 activation but alter caspofungin sensitivity, which requires Upc2. (A) Cells were treated with 1 mM SNP–0.5 mM SHAM (SS) for 1 h or 18 h. Activation of Mkc1 was determined using p44/42 antibody. Cells treated with 25 µg/ml calcofluor white (CFW) were used as a positive control. Activation of Hog1 was determined using p38 antibody, with cells treated with 2 mM hydrogen peroxide used as a positive control. The level of anti-actin on the respective blots is shown as a loading control. (B) Wild-type cells were grown with 1 mM SNP–0.5 mM SHAM and/or 100 ng/ml caspofungin (n = 3). (C) The upc2Δ mutant and the corresponding wild-type strain were grown in synthetic complete medium with 1 mM SNP–0.5 SHAM or in combination with 100 ng/ml caspofungin. A representative data set is displayed (n = 3). (D) Microdilution assays were performed with 3.9 ng/ml to 8 µg/ml caspofungin with or without 1 mM SNP–0.5 mM SHAM. The table shows the MICs for caspofungin for the wild-type strain and the upc2Δ mutant with or without SNP+SHAM (n = 3).