FIGURE 7.

Effect of the mini-TMD antibodies on GpHR deletion mutants. A, the ratio of RMFI for the mutant/WT was determined using the respective HinR-specific antibodies to estimate the relative expression of the mutant on cell surface and is designated as Re (24). Similar ratios were determined for the mini-TMD antibodies to determine the relative accessibility of the epitope in mutants and designated as Ra. An Ra/Re >1.2 indicates higher binding of the mini-TMD antibodies to the mutant as compared with the WT. Each bar represents RMFI ratios of three experimental replicates repeated three times. B, the WT and deletion mutants (HinTMD and TMD) of all the three GpHRs were transiently transfected, their basal cAMP production was measured in the presence of NRIgG or the mini-TMD antibodies (10 μg/ml) after 1 h of incubation, and cAMP produced was normalized to their cell surface expression (as determined in A). The statistical significance was compared with the respective pre-immune IgG control and is denoted by the p value calculated from the two-tailed unpaired t test.