Figure 5. The effects of JWH015 on Rac1 GTPase activity in trabecular meshwork cells

A: Time course of the effects of JWH015 on Rac1 GTPase activity in TM cells. Cells were serum-starved for 48 h, then stimulated with 100 nM of JWH015 for the indicated times. Rac1 GTPase activity in TM cells was assessed by PAK-PBD-GST pull-down assays. Blots shown are representative of three independent experiments. In the bar graphs, results are normalized to the density of the total Rac1 bands in the corresponding samples (n=3). B: The effect of SR144528 and SR141716A on JWH015-induced Rac1 GTPase activity in TM cells. TM 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 five min. The Rac1 GTPase activity was measured by PAK-PBD-GST pull-down assays. Blots shown are representative of three independent experiments. In the bar graphs, results are normalized to the density of the total Rac1 bands in the corresponding samples (n=3). JWH015 treatment led to a transient decrease in Rac1 activity. SR144528, but not SR141716A markedly inhibited the JWH015-induced inactivation of Rac1.