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Aspirin,the acetylsalicylic acid(ASA),is a most widely prescribed medication used to relieve pain,fever,and inflammation.Recent studies have revealed new benefits of aspirin,including reduction of heart attack or stroke,anti-cancer,and life-extension etc.Given profound effects of aspirin,the mechanism of action remains to be determined.Here,we used a deuterium-labeled aspirin together with mass spectrometry-based acetylome measurement,termed "DAcMS",to unravel the landscape of in vivo protein acetylation as induced by aspirin.We propose an in-depth atlas of aspirin-induced acetylome that reveals novel mechanistic insights into the biological roles of aspirin.