Localization of αi2 and sαi2 Expression vectors encoding αi2 (A and C) or sαi2 (B and D) were transiently transfected into COS-7 (A and B) or BHK (C and D) cells. Subcellular localization of the proteins was visualized by immunofluorescence microscopy using the EE monoclonal antibody followed by a Texas Red conjugated anti-mouse antibody, as described in "Materials and Methods". Bar, 10 μm. E, pcDNA3 (lanes 1, 2), EE-αi2-pcDNA3 (lanes 3, 4, 9, 10), EE-sαi2-pcDNA3 (lanes 5, 6, 11, 12) or EE-αqC9,10S-pcDNAI were transiently transfected into COS-7 (lanes 1–8) or BHK (lanes 9–12) cells. Cell lysates were separated into particulate (P) and soluble (S) fractions, as described in "Materials and Methods". EE-tagged Gα subunits were detected at 41–45 kDa (double asterisk) by immunoblotting with the EE monoclonal antibody. A slightly smaller protein is consistently detected (denoted by single asterisk) by the EE antibody, and this background band fractionates exclusively to the soluble fraction.
Wedegaertner BMC Cell Biology 2002 3:12 doi:10.1186/1471-2121-3-12