Figure 2
Effect of BAPTA-AM on [Ca2+]i and ERK1/2 activation. A: Measurement of [Ca2+]i. Hepatocytes were preincubated with 0.55 % DMSO or BAPTA-AM (40 μM) during the last 25 minutes of the fura-2 AM loading. After 60 seconds of registration the cells were stimulated with norepinephrine (10 μM) in the presence of timolol (10 μM). Results show a typical single cell response. B-D: ERK1/2 responses. Hepatocytes cultured for 3 h were pretreated for 30 min with BAPTA-AM (40 μM) in the presence of timolol (10 μM) prior to stimulation with norepinephrine (10 μM), A23187 (10 μM), TPA (1 μM) or PGF2α (10 μM) for 5 min before cells were harvested. All cultures contained DMSO at a concentration of 0.5 % during the preincubation and a final concentration of 1 % DMSO during incubation with agonist. B: Activity measurements of ERK1/2 representing mean ± S.E.M. of three experiments. C, D: Immunoblots using antibody against dually phosphorylated ERK1/2.