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For the last-month rule, the testing period begins with the last month of your tax year and ends on the last day of the 12th month following that month (for example, December 1, 2019, through December 31, 2020)

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In either situation, the cells were then permeabilized with cold PBST (0.5% Triton X-100) for 10 min, followed by fixation with 4% paraformaldehyde (PFA) in PBS (pH 7.4) for 15 min at room temperature

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