FIGURE 5.
Selective effects of hFFA1 binding pocket mutants on the function of lauric acid, GW9508X and rosiglitazone. A, receptor activation by rosiglitazone or GW9508X was measured in [35S]GTPγS incorporation assays performed on membranes prepared from HEK293 cells transfected with Gαq fusion proteins of wild type (WT) hFFA1 and H137A, R183A, N244A, R258A, and R183A,R258A receptor mutants (n = 2–7). B, wild type, H137A, R183A, N244A, R258A, and R183A,R258A hFFA1-eYFP were produced and used to generate Flp-In T-REx HEK293 cells. Each receptor construct was detected only following treatment of the cells with doxycycline. C, the effect of the above mutations was then tested in cell population calcium assays in response to lauric acid and rosiglitazone (n = 2–6). In A and C, the error bars indicate S.E.