Figure 6. The effects of JWH015 on phosphorylation of cofilin in trabecular meshwork cells

(A) Time course of the effects of JWH015 on phosphorylation of cofilin in TM cells. Cells were serum-starved for 48 h, and then treated with 100 nM JWH015 for the indicated times. TM cell lysates were prepared and subjected to western blot analysis using anti-phospho-cofilin and anti-cofilin antibodies. Blots shown are representative of three independent experiments. In the bar graphs, results are normalized to the density of total cofilin bands in the corresponding samples (n=3). (B) The effects of SR144528 and SR141716A on JWH015-induced phosphrylation of cofilin in TM cells. Cells were serum-starved for 48 h and pretreated with 1 μM SR144528 (SR2), 1 μM SR141716A (SR1), or vehicle for 30 min followed by stimulation with 100 nM of JWH015 for 30 min. TM cell lysates were prepared and subjected to western blot analysis using anti-phospho-cofilin and anti-cofilin antibodies. Blots shown are representative of three independent experiments. In the bar graphs, results are normalized to the density of total cofilin bands in the corresponding samples (n=3). JWH015 treatment decreased the level of phosphorylated cofilin in a time-dependent manner. Pretreatment with SR144528, but not SR141716A, markedly inhibited JWH015-induced dephosphorylation (activation) of cofilin.