FIGURE 4.

The effects of trypsin digestion of DIDS-treated rat erythrocyte ghosts on the cross-linked product of MCT1 with embigin. In panel A, rat erythrocytes were incubated in the absence or presence of 100 μm DIDS for 1 h at 37 °C prior to preparation of ghosts and, where required, treatment with trypsin as described under “Experimental Procedures.” The proteins were subjected to SDS-PAGE and Western blotting using the antibodies against the C terminus or 7/8 loop of MCT1 or an antibody raised against the cross-linked product of MCT1 with embigin (16). In B data are shown for samples that have been treated with N-glycanase F (DGaseF) as described under “Experimental Procedures.” The data shown are representative of two or three experiments that gave similar results.