Figure 5.

Biochemical verification of Kal7^KO. A, Total SDS lysates (20 μg protein) prepared from the parietal cortices and hippocampi of Wt, Het, and Kal7^KO mice were analyzed for Kal7 and βIII-tubulin. B, Levels of Kal7 and ΔKal7 in Kal7^+/KO mice were compared with levels in Wt mice. C, Parietal cortices from Wt and Kal7^KO mice were separated into crude subcellular fractions. Equal amounts of protein (5 μg) from each fraction were analyzed using Kal7-specific antibody (JH2959); similar results were obtained with affinity-purified Kal7-specific antibody (JH2958). The success of the fractionation was verified by visualizing synaptophysin and PSD-95. D, The splicing pattern for the region around the Kal7-specific exon is depicted; introns are not drawn to scale. Kal8 transcripts retain part of the adjacent intron (hatched), including a small protein coding region (gray) followed by a poly(A) addition signal and a poly(A) tract (Johnson et al., 2000). E, SDS lysates (20 μg) prepared from parietal cortices of Wt and Kal7^KO mice were visualized with a pan-Kalirin antibody, JH2582; the asterisk indicates a nonspecific band. F, Scans of gels were aligned, revealing the presence of Kal8 in Kal7^KO lysates; the asterisk indicates a nonspecific band. G, Levels of Kal12 and Kal9 in Het and KO mice were compared with Wt mice (n = 7). Error bars indicate SEM.