Figure 6.

The HOG pathway was activated by exposure to high iron levels. (A) Western blot analysis of phosphorylated Hog1p (P-Hog1p) in C. albicans SC5314 (WT) cells exposed to 0 or 30 μM FeCl3 in RPMI at 30°C for 10 min. 5 μg total protein per sample were separated by SDS-PAGE. Phosphorylated Hog1p was detected by exposure of the membrane for 100 sec (for P-Hog1p) and 130 seconds (for Hog1p) after HRP reaction. (B) Western blot analysis of phosphorylated Hog1p in C. albicans SC5314 cells exposed to 30 μM or 1.2 μM FeCl3 in YNB medium for 7.5, 10 or 15 min at 30°C. 16 μg total protein per sample were separated by SDS-PAGE. Phosphorylated Hog1p was detected by exposure of the membrane for 100 sec (for P-Hog1p) and 130 seconds (for Hog1p) after HRP reaction. (C) Western blot analysis of phosphorylated Hog1p (P-Hog1p) in C. albicans DAY286 cells exposed to 0 or 30 μM FeCl3 in RPMI at 30°C for 10 or 15 min. Sorbitol [1 M] was used as positive control. 12 μg total protein per sample were separated by SDS-PAGE. Phosphorylated Hog1p was detected by exposure of the membrane for 80 sec (for P-Hog1p) and 40 seconds (for Hog1p) after HRP reaction. (D) Western blot analysis of phosphorylated Hog1p in C. albicans DAY286 cells exposed to 30 μM or 1.2 μM FeCl3 in YNB medium for 0, 5, 10 or 20 min at 30°C. Procedures were the same as indicated above except the following: 16 μg protein per sample were loaded on the gel and the membrane was exposed for 20 sec (P-Hog1p) and 30 sec (Hog1p) respectively. The pictures were slightly rotated to obtain almost straight bands.

Kaba et al. BMC Microbiology 2013 13:16   doi:10.1186/1471-2180-13-16
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