Galleria mellonella infection was carried out as described previously [61 (link)] using a Hamilton syringe to deliver 106C. glabrata cells per waxworm. CFU analysis was done by cleaning the waxworm surface with 70% ethanol, puncturing it with a 1 ml syringe, and collecting the haemolymph from 25 worms per time point. Appropriate dilutions were made, plated on YPD agar plates, and incubated at 37°C for 24 h to count CFU.
Murine infections were carried out by intravenously injecting 6–8 week old female Swiss/Webster mice (Taconic) with 7.5 x 108 young or old C. glabrata cells isolated as described above. One group (n = 15) was injected with the antibody RB6-8C5 [93 (link)] every 48 h to deplete neutrophils. Depletion of neutrophils was verified by peripheral cell counts. Mice from both groups were sacrificed (n = 3) at 2 d and 4 d. The fungal burden was collected by homogenizing the kidneys to determine CFU on YPD plates, and budscar count by calcofluor staining (Sigma) as described previously [94 ].
Murine infections were carried out by intravenously injecting 6–8 week old female Swiss/Webster mice (Taconic) with 7.5 x 108 young or old C. glabrata cells isolated as described above. One group (n = 15) was injected with the antibody RB6-8C5 [93 (link)] every 48 h to deplete neutrophils. Depletion of neutrophils was verified by peripheral cell counts. Mice from both groups were sacrificed (n = 3) at 2 d and 4 d. The fungal burden was collected by homogenizing the kidneys to determine CFU on YPD plates, and budscar count by calcofluor staining (Sigma) as described previously [94 ].