Figure 7

Comparison of dose-response curves derived using CANDLES or cAMP ELISA. (A) An increasing stimulation of FSHR-293 (50,000 cells) with rFSH (0.01 mIU/ml to 1000 mIU/ml) also yields a dose-dependent increase in luminescent signal from co-cultured GS-293 (50,000 cells). The adjoining table shows the area under curve of luminescence values for different rFSH doses. (B) Luminescence values (RLU) in Figure 7A (20 min after stimulation) were used to calculate EC₅₀ for CANDLES (123.6 mIU/ml). A cAMP ELISA kit was used to compare EC₅₀ values with CANDLES. FSHR-293 cells (150,000 cells) stimulated with similar doses of rFSH (0.1 mIU/ml to 1000 mIU/ml) for 20 min were used to calculate cAMP values using ELISA. Data were normalized to percentage of maximal responses and curves were fitted using four-parameter logistic curve. EC₅₀ values calculated using CANDLES (123.6 mIU/ml) are very similar to those calculated using a cAMP ELISA kit (115.6 mIU/ml) and there is no statistical difference (p = 0.9885). (A and B) Data represented as mean of triplicates for one representative experiment (± SEM) with at least three independent repeats.