Skip to main content
. 2010 Jun;160(3):627–642. doi: 10.1111/j.1476-5381.2010.00681.x

Figure 7.

Figure 7

Interactions between CB1 receptor and β2-adrenoceptor pERK signalling in HTM cells. (A) Confocal immunofluorescence images of HTM cells using an anti-CB1 primary antibody with a FITC-conjugated secondary antibody (left panel), an anti-β2-adrenoceptor primary antibody with a Cy3-conjugated secondary antibody (middle panel) and the merged image (right panel). Scale bar is 20 µM. (B) Isoprenaline (ISO, 1 and 10 nM, 30 min) and forskolin (10 µM, 30 min) pCREB responses in HTM cells following 1 h DMSO or WIN (10 µM) pretreatment. **P < 0.01 compared with corresponding isoprenaline concentration pretreated with DMSO; n= 18–24. (C) pERK responses in HTM cells following a 10 min exposure to either aqueous vehicle and isoprenaline (100 nM) treatments, or to 0.05% DMSO vehicle and WIN (10 µM). Pretreatment with ICI (1 µM), CGP (1 µM), AM251 (1 µM) or AM630 (1 µM) was for 15 min and with PTx (100 ng·mL−1) for 24 h before isoprenaline or WIN application. PTx, ICI, CGP and AM251 bars are expressed as relative pERK level normalized to vehicle-treated cells that were also pretreated with the same antagonist or toxin to eliminate any affect these compounds may have had on the basal pERK levels. **P < 0.01 compared with vehicle, ***P < 0.001 compared with vehicle or DMSO; n= 8–27. (D) pERK dose–response in HTM cells following 10 min co-application of isoprenaline with 0.05% DMSO or 1 µM WIN; n= 13–16. (E) Isoprenaline pERK dose–response curves in HTM cells following 15 min pretreatment with either 0.05% DMSO or 1 µM AM251, followed by 5 min treatment with isoprenaline; n= 12–24. (F) Isoprenaline pERK dose–response curves in HTM cells following 15 min pretreatment with 0.05% DMSO or 1 µM O-2050 and 5 min exposure to isoprenaline; n= 12–16. AM251, N-(piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide; CREB, cyclic AMP response element binding protein; DMSO, dimethylsulphoxide; ERK, extracellular signal-regulated kinase; HTM, human trabecular meshwork; O-2050, (6aR,10aR)-3-(1-methanesulfonylamino-4-hexyn-6-yl)-6a,7,10,10a-tetrahydro-6,6,9-trimethyl-6H-dibenzo[b,d]pyran; PTx, Pertussis toxin; WIN, WIN 55,212-2.