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Proteins were transferred to nitrocellulose, total protein was assessed using FastGreen fluorescence labeling and Munc13-4 and STX7 were detected using anti-Munc13-4 (R&D, MAB89661) and anti-STX7 (R&D, AF5478) antibodies, respectively

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Indeed, some studies opted to use Rome III rather than Rome IV precisely because the latter is approximately 50% less sensitive in diagnosing IBS