Erythrocyte membrane protein analysis. Erythrocyte membranes were separated on 10% Laemmli gels and immunoblotted with the use of antibodies as shown. Loading: C1, C2, controls 1 and 2. P indicates proband [sdCHC(A)] except where labeled as glut1DS patient A or B. (A) The glut1 protein is heavily glycosylated (seen as a broad 50- to 100-kDa band). Scanning densitometry analysis of the deglycosylated glut1 band (labeled PNGase treated) showed normal amounts of glut1 in the patients with sdCHC and reduced amounts of glut1 (by ∼ 40%) in the patients with glut1DS. A protein 4.2 loading control is shown beneath each blot. The reduction in stomatin has been shown previously and is characteristic of sdCHC. Stomatin was present in normal amounts in the patients with glut1DS. (B) Both CD58 and Lutheran protein were increased in the sdCHC sample. An actin loading control is shown beneath each blot.