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. 2012 Jun 15;287(31):26365–26376. doi: 10.1074/jbc.M112.354696

FIGURE 4.

FIGURE 4.

TcSMUGΔGPI::GFP is secreted as a fully processed mucin. A, epimastigotes transfected with the indicated construct were incubated in serum-free medium and the conditioned medium (CM) and parasite pellet (P) fractions were probed with mAb 10D8. Signals provided by endogenous Gp35/50 mucins are indicated by a bracket. B, CM and P fractions from the indicated parasites were fractionated onto mAb anti-FLAG-Sepharose and retained fractions probed with rabbit antibodies to FLAG (left panel) or mAb 10D8 (right panel). C, suspensions of parasites expressing the indicated protein were labeled (+) or not (−) with Neu5Azα2–3LacβOMe (SA-az) followed by reaction of the azido group with a phosphine-tagged biotin, lysed, and probed with HRP-avidin by Western blot. Signals provided by endogenous Gp35/50 mucins are indicated by a bracket. D, lysates from the indicated parasites were labeled (+) or not (−) as in C, fractionated onto StreptAvidin-agarose and both flow-through (UB) and retained (B) fractions were probed with mAb anti-FLAG. Molecular markers (in kDa) are indicated.