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Intracellular ROS detection in S.pombe: Wild type yeast cells were grown till mid log phase in liquid rich YE media supplemented with adenine sulphate (100 μg/ml). Cells were treated with 2 mM of pyrogallol (from 100 mM stock solution prepared in water) for approximately 3 hours at 30^oC. H₂DCFDA dye was added at a final concentration of 100 μM concentration (stock concentration 10 mM in DMSO),  and cells were further incubated for 90 min at 30^oC to detect ROS by confocal microscope.

Yeast sporulation assay: S. pombe cells of opposite mating type were suspended in liquid media and 10 μl of the suspension was spotted on malt extract agar media containing different doses of pyrogallol. The plates were incubated for 3 days at 30^oC. The sporulation was monitored by staining the colonies with iodine vapour and also checked under the phase contrast microscope.

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