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Crg2 negatively regulates the intracellular cAMP level in C. neoformans. Cells were grown in YPD overnight and washed twice with distilled sterile water before being subjected to glucose starvation. Cell extraction and cAMP measurement were carried out as previously described (32). The assay was carried out in triplicate, and the mean values were plotted against time. The experiment was repeated twice with similar results. WT, wild type.
