FIG. 2.

PGE₂ production in C. albicans whole cells and cell-free lysates. (a) C. albicans strains SC5314 and CHN1 were grown in SDB at room temperature or 37°C with or without 500 μM AA and sampled at various times. Controls included no AA, AA alone, and HK Candida (boiled, 6 h). (b) For whole-cell prostaglandin production in serum, C. albicans strain SC5314 was incubated in normal or analbuminemic rat serum for 4 h at 37°C. (c) For Candida cell-free lysates, 24-h cultures of C. albicans strains SC5314 and CHN1 were lysed by vortexing with acid-washed glass beads in 1 mM EDTA buffer. Lysates were centrifuged, protein concentrations were normalized to a standard, and the lysates were incubated at 37°C with or without 500 μM AA for 90 min. Controls included buffer alone, buffer with AA alone, and lysates without AA. Both whole-cell supernatants and cell lysates were assayed for PGE₂ production using a monoclonal PGE₂ EIA kit (Cayman Chemicals, Ann Arbor, MI), while PGE₂ production in serum was measured using a PGE metabolite EIA kit (Cayman Chemicals, Ann Arbor, MI). Experiments were performed and measured in duplicate, and values are the averages of results of two to three independent experiments. *, P < 0.05, live versus HK C. albicans.