Fig. 6.

Pilot screen. (A) The LOPAC screening set (Library of Pharmacologically Active Compounds; Sigma Aldrich) containing ∼1280 compounds was tested at 10 μM against the Gal3/1ctR chimera in the cAMP biosensor assay under conditions as optimized in Figure 5 (10,000 cells per well), in the presence of an EC₉₀ of porcine galanin (antagonist mode). Compounds that displayed activity greater than three times the standard deviation from the mean of all the compounds tested were designated as hits (see Table 2). The S:B was 3.4 if signal was generated with an EC₉₀ of forskolin alone and baseline using an EC₉₀ of forskolin together with an EC₉₀ of galanin. An average Z score of 0.7 was calculated from eight wells of EC₉₀ forskolin alone (signal, open squares) and eight wells of EC₉₀ forskolin+EC₉₀ galanin (baseline, closed squares) on each plate (total n=32) and the primary hit rate was ∼5.4%. (B) Four of the top 10 hits were challenged at the indicated concentration ranges with an EC₉₀ of forskolin together with an EC₉₀ of galanin. The data presented in (B) are means±SEM of triplicate wells (n=3).