Figure 9. Hsp90 depletion affects MAP kinase signalling in the presence and absence of stress.

C. albicans wild-type and tetO-HSP90 cells (SN95 and CaLC1411: Table 1) were treated with 0 or 20 µg/ml doxycycline for seven hours. Mkc1, Cek1 and Hog1 phosphorylation levels were then assayed by western analysis in unstressed cells or cells treated as follows: (A) 30 minute 30°C–42°C heat shock; (B) 30 minutes with 100 µg/ml Calcofluor White (CFW); (C) 10 minutes with 5 mM H₂O₂; or (D) 12 minutes with 1 M NaCl. Total Mkc1 levels were assayed using Mkc1-6xHis-FLAG tagged cells in SN95 (CaLC681) and tetO-HSP90 (caLC648) cells (Table 1). Hsp90 levels and total kinase levels for Hog1 were also examined by western blotting relative to the internal Act1 loading control.