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Figure 9 | BMC Pharmacology

Figure 9

From: Delta Opioid activation of the Mitogen-activated protein kinase cascade does not require transphosphorylation of Receptor Tyrosine Kinases

Figure 9

Effect of antagonists to the insulin receptor (HNMPA-(AM)3) or the PDGF receptor (tyrosphostin-9) on ERK phosphorylation and tyrosine phosphorylation of the δ-OR by DSLET. C6-δ-OR cells were serum-starved overnight prior to exposure to DSLET (1 μM for 5 minutes). In some instances, DSLET exposure was performed in cells pre-treated with HNMPA-(AM)3 (100 μM) or tyrosphostin-9 (100 μM) 60 minutes prior to the δ-OR agonist. After agonist exposure, cell lysates were prepared and divided into aliquots for the analysis of ERK phosphorylation or tyrosine phosphorylation of the δ-OR. The δ-OR was immunoprecipitated as described in the methods section using the appropriate antibodies. Tyrosine phosphorylation of the δ-OR or ERK phosphorylation was determined by SDS PAGE, followed by immunoblotting of proteins onto nitrocellulose. Immunopositive bands corresponding to the tyrosine-phosphorylated form of the δ-OR (hatched bars) or ERK phosphorylation (open bars) were quantified using the MCID system. Band intensities are expressed graphically as: grain density × band area - [region of detection (ROD)] × pixel. Data points represent the mean ± SEM of three experiments. *p ≤ 0.05 compared to untreated samples as determined by ANOVA and the Tukey post-hoc test.

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