Moticam x camera

We collected 50–250 mL of water samples with detritus and/or plant material from ponds or lakes in Michigan in 2019–2021 (see Table S2 in the supplemental material). For Lake Erie, seston was collected by boat with a plankton net (≥20 µm) deployed 1–3 m from the surface, after which the collected material was transferred to a 50 mL conical centrifuge tube maintained at in situ water temperature in the dark. The samples were transferred to University of Michigan and incubated for ~1 month, at 20°C, under LED lighting. Water samples were observed using a Nikon TMS Inverted microscope (Nikon, Tokyo, Japan) to detect fungi associated with algae, micro-invertebrates, and protists. Detected fungal cells were photographed using Moticam X Camera (Motic, Hong Kong, China) or Dino-Eye Edge S Eyepiece Camera (AnMo Electronic Corporation, Taipei, Taiwan) digital cameras. Representative images were edited and assembled into plates using Adobe Photoshop. The cells were isolated manually using a manually prepared drawn-out glass capillary pipette. The isolated cells were washed by serial transfer in small drops (more than five) of UV-sterilized water, transferred into 200 µL PCR tubes with 1–2 µL of water, and kept at −80°C until DNA extraction.

Cells were transfected in the same manner as the clonal survival assays. At 48 h after transfection, cells were collected using trypsin, counted, and plated in a final volume of 200 µL/well of complete media with 10% FBS in a 96-well Ultra Low Attachment plate (1500 cells/well for WM1366; 600 cells/well for A375; Corning, Tewsbury, MA, USA). The plate was spun in a centrifuge with a plate rotor for 5 min at 200× g. At 96 h after plating, pictures were captured of the different conditions with transmitted light microscopy (Moticam X camera, Motic, Richmond, BC, Canada and an Axiovert microscope on the 5× objective, Zeiss, Thornwood, NY, USA).