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. 2003 Oct 1;140(3):547–557. doi: 10.1038/sj.bjp.0705464

Figure 1.

Figure 1

Time course of THC-induced activation of JNK. Cortical neurons were exposed to THC for 5–120 min, then cells were harvested and the cytosolic fractions analysed for the expression levels of the phosphorylated active forms of JNK 1 and JNK2 using Western immunoblot. (a) A significant increase in phospho-JNK1 expression was found following treatment with THC (5 μM) for 5 min. Exposure to THC for 30 and 60 min had no effect on phospho-JNK1 expression. However, treatment with THC for 120 min significantly decreased phospho-JNK1 expression. Results are expressed as mean±s.e.m. for seven observations, ***P<0.001, *P<0.05. (b) Phospho-JNK2 expression was significantly increased when neurons were treated with THC (5 μM) for 120 min. Exposure to THC for 5, 30 and 60 min had no effect on phospho-JNK2 expression. Results are expressed as mean±s.e.m. for seven observations, ***P<0.001. (c) A sample Western immunoblot demonstrating the increase in phospho-JNK1 expression at 5 min of THC treatment and the corresponding increase in phospho-JNK2 expression following treatment with THC for 120 min. (d) Neurons were treated with THC (5 μM) for 5 min, then cells were harvested in lysis buffer and the cytosolic fractions analysed for total JNK1 protein expression using Western immunoblot. THC had no effect on JNK1 protein expression at 5 min. Inset: A sample Western immunoblot showing that THC treatment for 5 min had no effect on JNK1 protein expression as compared to vehicle-treated neurons. (e) Cortical neurons were treated with THC (5 μM) for 2 h, then cells were harvested and analysed for total JNK2 protein expression by Western blot analysis. Incubation of cells with THC for 2 h did not affect JNK2 protein expression. Inset: A sample Western immunoblot demonstrating total JNK2 protein expression in vehicle-treated neurons and neurons treated with THC for 2 h. (f) Untreated cortical neurons were harvested and analysed for phosphorylated JNK1, JNK2 and JNK3 expression. JNK3 expression was moderate compared to JNK1/2 expression. (g) A sample Western immunoblot showing that a 5 min treatment with THC increases phospho-JNK1 expression in a dose-dependent manner. Similar results were observed in five separate experiments. (h) A sample Western immunoblot showing that a 120 min treatment with THC increases phospho-JNK2 expression in a dose-dependent manner. Similar results were observed in five separate experiments.