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Briefly, brain slices were air-dried at 39C for 2 hours, washed with 1 PBS, treated with 3% hydrogen peroxide in methanol and Target Retrieval buffer (Advanced Cell Diagnostics [ACD]) for 15 minutes, and subjected to RNAscope Proteinase III incubation at 40C for 15 minutes

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Statistical comparisons between means were performed using an unpaired Students t-test to assess differences between two groups

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Pathophysiology The pathophysiology of CVS is yet to be established although several potential underlying mechanisms have been postulated