Orca c4742 95 12er

For fluorescence microscopy to visualise GFP- and DsRed-expressing cells, transformants were cultured in SR medium for 18 hours at 30 °C, then, cells were suspended into SG medium and incubated for 4-6 hours for GAL1 promoter induction. Finally, cells were harvested by centrifugation and observed directly. Cells were examined under Eclipse TE2000U microscope (Nikon, Tokyo, Japan) and digital images were acquired with an Orca C4742-95-12ER charge-coupled-device camera (Hamamatsu Photonics, Hamamatsu City, Japan) and processed by HC Image (Hamamatsu).

For the in vivo fluorescence microscopy of cells expressing pJMCS-DM23 (GFP-2XPH-PLCδ1) [36 (link)], exponentially growing SR-G cultures treated or not with neomycin 5 mg/mL were collected by centrifugation at 5000 rpm for 1 min and directly observed with an Eclipse TE2000U microscope (Nikon, Tokyo, Japan) using the appropriate sets of filters. For pRS410-GFP-LactC2 observation, the same procedure was followed but cells were cultured in SD. Digital images were acquired with an Orca C4742-95-12ER charge-coupled device camera (Hamamatsu, Hamamatsu city, Japan) and processed with HCImage software (Hamamatsu). Statistics on cell populations were performed by counting >100 cells for each experiment.