Figure 8.

PSD preparations from Wt and Kal7^KO mice. A, Purified ΔKal7 was used to quantify the amount of Kal7 in the average 1.1 GDa PSD (Collins et al., 2006; Sheng and Hoogenraad, 2007). Recombinant purified ΔKal7 was quantified by making a dilution series and comparing Coomassie Blue staining to a bovine serum albumin standard of known concentration. Western blot signals, using affinity-purified Kal7 antibody, for the ΔKal7 standard were used to construct a standard curve to quantify Kal7 in the PSD samples. B, Use of the pan-Kalirin antibody revealed that PSDs (5 μg of protein) prepared from Kal7^KO mice had increased levels of Kal9 and Kal12 compared with PSDs prepared from Wt mice. C, Left, PSDs (5 μg of protein) purified from the cortices of two sets of Wt and Kal7^KO mice were blotted for AMPA and NMDA receptor subunits, neuroligins 1 and 2, δ-catenin, and for the Kal7 interactors PSD-95, Rac1, and PP1. Right, Both the Triton-soluble (S) and insoluble PSD (P) fractions were blotted for NR2B and Cdk5. D, Data from three sets of Wt and Kal7^KO PSDs were averaged (n = 2 for Cdk5 TX S); *p < 0.05; **p < 0.01. Error bars indicate SEM.