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Fig. 5. Solubility properties of wild-type and mutant GFAPs produced in SW13Vim– cells and extracted with Triton X-100 buffer with increasing salt concentrations. Transfected SW13Vim– cells were extracted with a 0.5% Triton X-100 buffer without KCl (A,B) or with buffer also containing either 0.5 M KCl (C,D) or 1.0 M KCl (E,F). Nitrocellulose transfers were probed with an anti-GFAP antibody (Chemicon) (A,C,E) and then stripped and reprobed with an anti-actin antibody (B,D,F). Lanes: S, supernatant fraction; P, pellet fraction after centrifugation at 16,000 g; untxft, untransfected cells; wt, wild-type GFAP; mut, mutant GFAP; c, Triton-X-100-insoluble fraction from primary rat astrocytes, as positive control. Mr, molecular weight markers (60 kDa and 40 kDa).





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