Figure 5. Heat killed Candida albicans stimulates ERK1/2, p38 and NFκB.

(A) BMDMs were stimulated for the indicated times with 10⁶ cells/ml heat killed C. albicans. Cells were lsyed in SDS sample buffer and the levels of phospho ERK1/2, total ERK1/2, phospho p38, total p38, phospho JNK, phospho p105 and total IκB were determined by immunoblotting. (B) BMDMs were incubated with 2 µM PD184352 or 5 µM SB203580 as indicated for 60 min prior to stimulation with 10⁶ cells/ml heat killed C. albicans for 60 min. Cells were lsyed in SDS sample buffer and the levels of phospho ERK1/2, total ERK1/2, phospho p38, total p38 and phospho MK2 were determined by immunoblotting. (C) BMDMs were stimulated with 10⁶ cells/ml heat killed C. albicans. Total RNA was isolated and the levels of IκBα mRNA determined by Q-PCR. Error bars represent the standard deviation from 4 independent cultures. (D) Where indicated BMDMs were treated with 15 µM BMS 345541 or 10 µM BAY 65-1942 for 1 h. Cells were then stimulated with 10⁶ cells/ml heat killed C. albicans for 4 h, and IκB levels determined by Q-PCR. Error bars represent the standard deviation from 4 independent cultures.