Figure 5.

The SCSD-associated SPINT2 mutants are expressed as lightly glycosylated HAI-2 in monomer and multiple complex forms. Expression of wild type and mutant HAI-2 proteins was induced by treatment with different concentrations of doxycycline (Dox.), as indicated, in HAI-2 knockout Caco-2 variants engineered to express wild-type HAI-2 (A) or one of the three SCSD-associated mutants, R148C HAI-2 (B), R148H HAI-2 (C), and Y163C HAI-2 (D). Equal amounts of lysate protein was analyzed by immunoblot under non-reducing and non-boiled conditions for lightly glycosylated HAI-2 species using the HAI-2 mAb XY9. HAI-2 monomer and its oligomer forms (see fig. 6) were indicated.