Figure 4

The LHCGR extracellular domain (ECD) expressed in HEK-293 specifically reacted with human LHCGR mouse monoclonal antibody, LHR29. HEK 293 cells (expressing human LHCGR ECD, amino acid residues 1–362) were grown exponentially in tetracycline-free fetal bovine sera in the absence (lanes 1 and 3 in a and b, respectively) and in the presence of tetracycline (lanes 2 and 4 in a and b, respectively). Each lane contains 25 μg of total protein separated via electrophoresis through 8% polyacrlamide SDS gels. Blots were immuno-reacted with antibodies (lanes 3 and 4 of a and b, respectively). A shorter exposure of lane 4 (a) is shown on the right hand side of the Fig. 4a. Following chemiluminescence detection, blots were stained with coomassie brilliant blue (lanes 1 and 2 of a and b, respectively).