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. 2011 Aug 10;10(12):M110.007369. doi: 10.1074/mcp.M110.007369

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© 2011 by The American Society for Biochemistry and Molecular Biology, Inc.

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Fig. 5. — Validation of metacaspase-3 as a SUMOylation target in vivo. A stable cell line of epimastigotes expressing T. cruzi metacaspase-3 tagged with a 3 × FLAG (TcMCA3–3×FLAG) was grown in the presence of tetracycline for 60 h before cell lysate was prepared. TcMCA3–3×FLAG protein was purified by immunoaffinity chromatography using anti-FLAG M2-agarose affinity gel. Eluate was electrophoresed on 7.5% SDS-PAGE, transferred to nitrocellulose membranes and probed with either anti-FLAG (left) or anti-SUMO (right) antibodies. Protein bands corresponding to metacaspase and SUMOylated metacaspase-3 are shown with an empty or filled arrowhead.