Figure 4

Validation of the cell-based hit compounds. A. Force inhibition of pre-contracted ASM tissues from inherently hyper-responsive Fischer rats. Tracheal rings were first contracted for 10 min with acetylcholine (3 μM) and subsequently treated with increasing concentrations of compound 85070. For control, we used 5% w/v cyclodextrin. Data are presented as mean ± SE (n = 4 separate experiments). B. Stiffness responses of smooth muscle cells isolated from aorta and trachealis of the inherently hyper-responsive Fischer rats. Cells were treated with vehicle control (0.5% w/v cyclodextrin), dibutyryl-cAMP (1 mM), or compound 85070 (20 μM, 50 μM or 200 μM). Stiffness changes are normalized to respective baseline stiffness of an individual cell. Data are presented by geometric means ± SE (n = 127 to 505 cells). For each treatment, * indicates P < 0.001 and # indicates P < 0.05 between the cell types. For each cell type, ** indicates P < 0.001 when compared with respective vehicle control.