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. 1999 Nov 1;19(21):9469–9479. doi: 10.1523/JNEUROSCI.19-21-09469.1999

Fig. 3.

Fig. 3.

CALI of NCAM-180 intracellular domain disrupts brain spectrin binding in vitro. Purified chick NCAM (all isoforms) was incubated with purified bovine brain spectrin after CALI using 4d, 5e, or nonspecific IgG. The complex was immunoprecipitated with anti-NCAM, dissociated in Laemmli sample buffer, fractionated by SDS-PAGE, and immunoblotted with anti-spectrin antibodies to assay NCAM-180/spectrin binding. A shows spectrin immunoblot, whereas B shows the same filter after being stripped and probed with anti-NCAM (all isoforms), showing that there were equivalent amounts of NCAM present in each lane.Lane 1, Anti-NCAM immunoprecipitation; lane 2, no anti-NCAM control; lane 3, preincubation of NCAM with MG-labeled nonspecific antibody (IgG); lane 4, the same conditions as lane 3 except subjected to laser irradiation; lane 5, preincubation with MG-labeled 4d (intracellular domain of NCAM-180); lane 6, same conditions as lane 5 except subjected to laser irradiation (CALI using 4d); lane 7, preincubation with MG-labeled 5e (extracellular domain of all NCAM isoforms); lane 8, same conditions as lane 7, except subjected to laser irradiation (CALI using 5e).