Fig. 6.

cAMP levels are reduced in PN/PH cells treated with ACY-1215: A: PN/PH confluent cells were treated with ACY-1215 (10 µM) or DMSO for 16 h. Note that cAMP levels are higher in PN cells than in PH cells and are reduced by ACY-1215, particularly in the case of the PN cells. B: PN/PH confluent cells were treated with ACY-1215 (10 µM) or DMSO for 16 h and then treated with adenylyl cyclase activator forskolin (100 µM) or the phosphodiesterase inhibitor isobutyl methylxanthine (IBMX, 100 µM) for 30 min before the cells were harvested for assay. Note that ACY-1215 reduces both resting cAMP levels and forskolin-dependent increases in cAMP. cAMP levels were measured with a direct cAMP Enzyme Immunoassay Kit (no. CA200; Sigma) based on the manufacturer’s protocol. Results are expressed as pmol/ml. Bars represent means ± SE. *P < 0.05, **** P < 0.0001, and ####P < 0.0001. Statistical analysis was performed using a 2-tailed Student’s t-test. Each set of data is from 6 individual wells for A and 5–10 wells for B. *Compared with PN cells. #Compared with PH cells.