Fig. 10.

The effect of Ang II on MAP kinase activity in neuronal cultures. A, Neuronal cultures were incubated with 100 nm Ang II for indicated time periods, cell extracts were prepared in lysis buffer, and MAP kinase was immunoprecipitated by specific antibody (ERK-2). The resulting immunoprecipitates were electrophoresed on a 10% SDS/PAGE containing 0.5 mg/ml myelin basic protein. This was followed by an in-gel assay for MAP kinase as described in Materials and Methods. Top, Representative autoradiogram. Bottom, Data from three in-gel assays were presented as mean ± SE. Asterisks indicate significantly different from zero time control (p < 0.05). B, Neuronal cultures were incubated without or with 100 nm Ang II for 10 min in the absence or presence of 10 μm losartan (Los.) or 10 μm PD123319 (PD) as indicated. MAP kinase activity was determined as described in the legend to Figure 9.Top, Representative autoradiogram. Bottom, Data from three in-gel assays presented as mean ± SE. Asterisksindicate significantly different from untreated control (p < 0.05); double asterisk indicates significantly different from Ang II-treated neurons.