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Fluorescence staining using TOTO-1 (514/533; Invitrogen, Carlsbad, CA) was performed to confirm the damage to spore membranes caused by the antifungal substance. The spores were collected as described above and resuspended in 300 µL diluted TOTO-1 (1/5000 with TE buffer) for 3 min. big slot toto
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