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Cells were cultivated at 30°C in YPD medium (1% yeast extract, 2% bacto peptone and 2% glucose) to a log phase density of 1 × 10 cells/ml (sample A collection), then and diluted to 5 × 10 cells/ml and subjected to stress by adding a equal volume of 30°C pre-warmed YPD medium containing 2M KCl.
After the incubation with the cysteine solution, the plate was washed with 50 μL of PBS-T three times and blocked with 50 μL of 1× blocking buffer (prepared by diluting 2× blocking buffer with a equal volume of PBS-T) for 1 h at room temperature with gentle shaking.
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As a control, an equal volume of PBS was likewise injected.
As a control an equal volume of 2%PVP400 was likewise injected.
As a control, an equal volume of normal saline (200 µL/injection) was used.
As a control, a similar tube had an equal volume of buffer added to it.
A control used an equal volume of buffer instead of enzyme solution.
The upper aqueous phase was transferred in a tube containing an equal volume of isopropanol.
So mix up a teaspoonful of salt with an equal volume of vinegar in a cup.
The other group received a sham injection of an equal volume of the buffer.
Afterwards, an equal volume of buffer A containing 2% Triton X-100 was added.
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