Fig. 2

Construction and expression of Gα_i1 chimeras. A Homologous replacements or point mutations on putative effector-interacting domains were made between Gα_i1 (black) and Gα_t1 (orange) (Chi1-4) or Gα_q (green) (Chi5 and Chi6). Sites of replacement/mutation are indicated by their residue numbers. The locations of GTPase-deficient mutations, namely R178C and Q204L, and PTX-insensitive mutation (C351I) are highlighted with yellow dots. B Expression of the chimeras was verified by Western blotting. HEK293 cells in a 24-well plate were transfected with 0.2 μg of various chimeric constructs and the cell lysates were subjected to immunoblotting using antibodies against Gα_i1 and β-actin. Expressions of the chimeras were compared with that of Gα_i1