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. 2005 Jan 26;25(4):869–879. doi: 10.1523/JNEUROSCI.3212-04.2005

Figure 2.

Figure 2.

Similar spatiotemporal expression patterns and complex formation of IRSp53 and PSD-95 in the brain. A, Expression of IRSp53 in different brain regions. Membranes from various regions of adult rat brain were immunoblotted for IRSp53 and β-tubulin (control). The widespread expression of IRSp53 is similar to that of PSD-95 (Cho et al., 1992). Cb, Cerebellum; Ctx, cortex; Hc, hippocampus; St, striatum; R, the rest of the brain. B, Similar developmental expression patterns of IRSp53 and PSD-95. Membranes of rat brain at the indicated developmental stages were immunoblotted for IRSp53, PSD-95, and β-tubulin (control). E, Embryonic day; P, postnatal day; wk, week. C, Cofractionation of IRSp53 and PSD-95 in subcellular fractions of adult rat brain. Synaptophysin (SynPhy) was probed as a control. H, Homogenates; P2, crude synaptosomes; S2, supernatant after P2 precipitation; S3, cytosol; P3, light membranes; LP1, synaptosomal membranes; LS2, synaptosomal cytosol; LP2, synaptic vesicle-enriched fraction. D, Coenrichment of IRSp53 and PSD-95 in PSD fractions. PSD fractions of adult rat brain extracted with Triton X-100 once (PSD I), twice (PSD II), or with Triton X-100 and the strong detergent Sarcosyl (PSD III) were immunoblotted with IRSp53, PSD-95, and synaptophysin antibodies. E, DOC extracts of the crude synaptosomal fraction of adult rat brain were immunoprecipitated with IRSp53 antibodies or rabbit IgG (negative control) and immunoblotted with the indicated antibodies. Rb, Rabbit. F-J, The same extracts were immunoprecipitated with PSD-95, chapsyn-110, SAP97, SAP102, and S-SCAM antibodies or control IgGs and immunoblotted with the indicated antibodies. NR2B, NMDA glutamate receptor subunit 2 (positive control); Gp, guinea pig.