Effect of IP3 receptor antagonist on acinar cell function with or without nicotine Primary cells were washed and incubated at 37 C without or with 80 mM 2 APB for 30 minutes, fol lowed by an additional 6 minutes incubation with nico tine. Amylase released into the incubation medium was measured with Procion yellow starch as substrate. The data are presented as % initial trichostatin a mechanism of action content and represented as the meanSEM of four experiments. Effect of ERK Inhibitor on primary cell function with or without nicotine To study the effect of an ERK12 inhibitor on basal and stimulated amylase release, primary cells were pretreated with UO126 for 30 minutes. After washing, the cells were incubated with 100 uM nicotine for 6 min utes.

At the end of the incubation period, the cells were washed with HR buffer, and incubated with either buffer, or buffer containing 10 9 M CCK 8, for 30 minutes. Amylase release in the media and protein concentration were determined as outlined in the earlier section. The method used for CCK stimulation was similar Inhibitors,Modulators,Libraries to that described in the earlier section. Effect of JNK inhibitor or p38 inhibitor on primary cell function, with or without nicotine Primary cells were washed and incubated at 37 C with or without 100 nM JNK inhibitor or with 1 uM p38 Inhibitors,Modulators,Libraries kinase inhibitor for 30 minutes, followed by an add itional incubation with nicotine for 6 minutes. Cells were washed and then incubated at 37 C with or without CCK 8 for 30 minutes. Amylase released into the incu bation medium was measured with Procion yellow starch as substrate.

The data are presented as % initial content and represented as the meanSEM of four experiments. Statistics Results were reported as meanstandard error of the mean. The results were analyzed by students t test and where required, Inhibitors,Modulators,Libraries with analysis of variance. Differences were considered significant at a probability of 0. 05 or lower. Results Nicotine Effects on CCK stimulated acinar cell function The time and nicotine dose effects on maximal stimu lated amylase release in response to CCK are shown in Figure 1. The solid line represents the amylase response of control cells while the broken line represents the sti mulated amylase response by CCK at a maximal stimu lating dose of 10 9 M. The bottom panel in Figure 1 shows that nicotine at a dose of 100 uM induced max imal release of amylase while Inhibitors,Modulators,Libraries the upper panel shows that the maximal response occurring at 3 mins of exposure that persisted until 6 min before de cline.

Based on this observation, a single dose of nicotine and time of exposure for 6 min was used in subsequent experiments Inhibitors,Modulators,Libraries on the results as shown below. Effect of mecamylamine or conotoxin on primary cell http://www.selleckchem.com/products/Perifosine.html function, with or without nicotine The effects of nicotine in the presence or absence of mecmylamine, and conotoxin on the basal and cell sti mulated primary acinar cell function are shown in Figure 2.