Figure 5.

Effect of calpain inhibition on time course of loss of surface NR2B. A, Top panel, Representative Western blots probed with NR2B-specific antibodies after isolation of biotinylated surface proteins (“Biot”) and unbiotinylated internal proteins (“Sup”) as in Figure 4A. Shown is NR2B in WT and YAC128 MSN cultures at 0, 12 and 24 h following treatment of live MSNs with biotinylating reagent. Bottom panel, Plot shows time course of loss of biotinylated, full-length NR2B over 24 h in untreated WT and YAC128 9 DIV MSN cultures. n = 4 independent experiments each from different batches of WT and YAC128 MSN cultures. Difference is significant for time (p < 0.001), genotype (p < 0.001), and interaction between time and genotype (p < 0.01) by two-way ANOVA; **p < 0.01, ***p < 0.001 by Bonferroni posttests. B, Top panel, Representative Western blots probed with NR2B-specific antibodies as in A, except that MSN cultures were pretreated with CI-1 (1 μm) for 1 h before incubation with the biotinylating reagent, and then 1 μm CI-1 was maintained in the medium for the duration of the experiment. Bottom panel, Plot shows time course of loss of biotinylated, full-length NR2B over 24 h in CI-1-treated WT and YAC128 9 DIV MSN cultures. N = 3 and 4 independent experiments from different batches of WT and YAC128 MSN cultures, respectively. No significant difference between genotypes by two-way ANOVA.