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PA-TU-8902, MIA PaCa-2, HEK293T and U2OS cells were cultured in Dulbeccos Modified Eagles medium (DMEM, P04-03550, PAN Biotech) supplemented with 10% FBS, 1% NEAA and 1 mM sodium pyruvate (P04-43100, PAN Biotech)

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The thiamine-induced increases in contractions of the intestine smooth muscle and peristalsis, observed in a number of independent in vitro experiments on muscle strips of gastrointestinal tract (207, 219222), support the action of thiamine on ACh signaling in ENS in the absence of central regulation (236)