Figure 4.

Lack of association of lectin–GFP fusion protein 224 with endogenous lectin subunits. Amebic proteins were separated under nonreducing conditions by SDS-PAGE and transferred to PVDF membranes. The blot was cut into three equivalent blots and each blot was incubated with antibody to either a lectin-heavy subunit, lectin-light subunit, or GFP. The blots were then incubated in alkaline phosphatase-conjugated secondary antibody and developed in substrate for alkaline phosphatase. The position of the molecular mass markers (in kDa) is shown. 1, amebae expressing luciferase as a control; 2 and 3, independent amebae transfectants expressing the 224 fusion protein.