Figure 6. Lack of increase in NR2B phosphorylation in EphB1 KO mice following formalin or carrageenan injection.

Tyrosine phosphorylation of the NR2B subunit of the NMDA receptor was analysed by immunoprecipitation followed by Western Blotting in WT and EphB1 KO mice after injection with formalin or carrageenan. Saline injected animals were used as controls. Spinal cords were dissected from injected animals 1 h (A) or 3 hours (B) after injection. Representative immunoblots are shown in the top part of panels A and B. In panel A, the last two columns represent positive (mouse brain tissue) and negative (immunoprecipitation of mouse brain tissue conducted in the absence of anti-NR2B antibodies) controls of the immunoprecipitation and Western blotting protocol. The blots were quantified, and the ratio of phosphoTyr to the total NR2B present was calculated. The bar charts in the bottom half of panels A and B show the mean and SEM of the levels of phosphoNR2B present in EphB1 KO and WT mice following injection with Formalin, Carrageenan or saline, as indicated. There was a significant increase in Tyrosine phosphorylation of NR2B in WT mice following formalin (A) and carrageenan injection (B), p<0.05. There was no change in the level of NR2B phosphorylation in EphB1 KO mice following either injection. Comparisons performed with t-test for WT in the formalin experiment and EphB1 KO in the carrageenan experiments, with Mann-Whitney U-test in the remaining experiments. * p<0.05 vs respective saline. For all experimental groups n = 3, except in A) EphB1 KO saline n = 4.