Fig. 2

Calcineurin function is essential for cell survival and integrity of C. albicans cells lacking both CaGDT1 and CaPMR1. Quantification of clonogenic survival of cells treated with CsA for different time (a) and different concentrations (b) as indicated. Clonogenic survival was determined by plating cells onto YPD plates, and colony-forming unit (CFU) was counted manually. Data are means of three independent experiments. Empty bar indicates the colony-forming unit (CFU) of the cell culture immediately before the CsA treatment. Black bars and grey bars represent CFUs of cell cultures not treated and treated with 25 μg ml^− 1 CsA, respectively. Morphology of cells untreated (c) and treated with CsA for 8 h (d). e Cell cultures untreated (left test tube) and treated with CsA for 8 h (right test tube). Representative epifluorescent micrographs of cells untreated (f) and treated with CsA for 8 h (g) after being co-stained with both Annexin V-FITC and propidium iodide. Epifluorescent micrographs of cells untreated (h) and treated with CsA for 4 h (i), after being incubated with the vacuole indicator dye FM4–64 for 30 min. The wild-type (WT) and the double-gene gdt1/gdt1 pmr1/pmr1 mutant (DM) were cultured at 30 °C in YPD to log phase before they were treated with CsA of 25 μg/ml for these experiments. Cells with normal cell morphology but without intracellular FM4–64 staining are indicated with arrows, while cells with abnormal internal structures are circled