FIG 4.

Mutants' alterations to exo-GXM release correlate with adherence. A total of 10⁶ C. neoformans cells was seeded into individual wells of 96-well polystyrene plates and incubated at 37°C. After 48 h, the wells were washed to remove nonadhered and/or weakly adhered cells before resuspension in XTT for colorimetric analysis of metabolic activity as a proxy for viable cell count. (A) OD₄₉₀ readings from cells grown in YNB medium, normalized to wild-type cell readings. liv7Δ mutant 1/2 cell adherence was reduced, and ima1Δ mutant 1/2 cell adherence was increased compared to that of wild-type cells. (B) OD₄₉₀ readings from cells grown in 10% Sabouraud's medium, pH 7.3, normalized to wild-type cell readings. ima1Δ mutant 1/2 cell adherence was increased compared to that of wild-type cells. Combined data from three independent experiments are shown. P values were calculated using a Mann-Whitney test; bars represent means with standard deviations. (C) OD₄₉₀ readings from cells grown in YNB medium, normalized to wild-type cell readings. Cells were mixed with purified GXM or vehicle (PBS) prior to inoculation of plates. Wild-type and liv7Δ mutant 1/2 cell adherence increased in the presence of GXM.