Fungal Biofilm Formation Assay

Biofilm assays were performed based on previously published protocols 14 (link)52 (link), with minor modifications. Briefly, overnight fungal cultures were washed three times in PBS and resuspended in Dulbecco's modified eagle medium (DMEM, Gibco) at a final concentration of 10⁶ cells ml^-1. For some experiments, DMEM was supplemented with 1 mM, 50 mM, or 100 mM LiCl, 100 mM NaCl, 20 µg ml^-1 EBS, or a combination of these chemicals. Next, 200 µl of fungal cells were transferred into individual wells of sterile, polystyrene, flat-bottom, 96-well microtiter plates (Corning). Wells with medium only were included as controls. To minimize evaporation, 200 µl sterile water was added to the wells surrounding the test wells. Plates were incubated at 37°C and 5% CO₂ for 48 h.