Figure 4

Phosphorylation of threonine and tyrosine residues on spinal cord NR1 subunits. No phosphorylation of NR1 threonine or tyrosine residues was found in response to peripheral inflammation. A. Lumbar spinal cord NR1 subunits were immunoprecipitated with an antibody that recognizes all NR1 splice variants or an antibody that recognizes splice variants with the C1 cassette before and after left hind paw inflammation with carrageenan. The protein was run on western blots and probed with an anti-phosphothreonine antibody. The western blots at the top are representative experiments and the graph at the bottom presents the mean ± SEM band density of all experiments (Total NR1; N = 8 rats per time point, C1 cassette; N = 9 rats per time point). ANOVA p > 0.05. The molecular weight of the bands on the western blots was ~118 kd. B. NR1 subunit protein was immunoprecipitated with an antibody that recognizes all NR1 splice variants or an antibody that recognizes splice variants with the C1 cassette, run on western blots and probed with an anti-phosphotyrosine antibody. The western blots at the top are representative experiments and the graph at the bottom presents the mean ± SEM band density of all experiments (Total NR1; N = 4 rats per time point, C1 cassette; N = 4 rats per time point). ANOVA p > 0.05.