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For experiments proceeding over 21 days, 3 ml of the expended medium were exchanged with 5 ml of fresh HPC medium on day 7 and another 5 ml were exchanged on day 14.
Food and medium were exchanged daily.
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The medium was exchanged with fresh medium lacking phenol red.
Twenty-four hours later, medium was exchanged against 100 μl of medium containing different concentrations of verteporfin (0.001 10 μM) was added.
Culture medium was exchanged after 5 days in Space and the cells were fixed after 10 days.
30 min before the start of recording, the culture medium was exchanged to fresh warm medium (300 μl/well) to remove floating debris.
Every 7 days, 50% of the supplemented BrainPhys medium was exchanged and volume loss due to evaporation was compensated with dH2O (CRTD media kitchen).
Then, the medium was exchanged into 250 µL of DMEM and yeasts were additionally cultivated for 12 h at 30°C.
The medium was exchanged daily.
After 4 h, culture medium was exchanged.
Medium was exchanged twice before fluorescence measurements.
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