FIGURE 8.
Formation of chicken-rat AChE hybrid dimers. A, HEK293T cells were single or double transfected with cDNAs encoding rAChET and cAChET. Sucrose density gradient analysis was performed as in Fig. 1A. Enzymatic activities are expressed in arbitrary units, and one representative result is shown (n = 3). B, the cell lysates were immunoprecipitated by anti-cAChE or anti-rAChE antibodies and analyzed by SDS-PAGE and Western blotting with anti-cAChE or anti-rAChE antibodies. C, extracts from cells expressing rAChET and cAChET separately or together were analyzed by nonreducing SDS-PAGE and labeled with anti-cAChE and anti-rAChE antibodies. The presence of an intermediate band between cAChE and rAChE dimers, recognized by both antibodies, demonstrates the formation of cAChET-rAChET hybrid dimer. Representative gels are shown (n = 3). Ab, antibody; IP, immunoprecipitation; IB, immunoblot.