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The four cerebellar tissue slices were then laid onto a Millicell-CM culture insert (Millipore, 0.4 m) and maintained in slice culture medium (50% MEM with Earle's salts, 35% Earle's balanced salt solution, and 15% heat-inactivated HS) at 37 C for 3 days

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For unmodified sucA, binding data was collected at the highest concentration of 500 nM