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Fig. 7. Expression profile and subcellular localisation of SIQ1098A and SIAla3Q in COS-1 cells. (A) The cDNAs of the SIQ1098A and the Ala triplet AlaNDQ mutants were transfected into COS-1. The cells were biosynthetically labelled for 4 hours with [35S]methionine, lysed and immunoprecipitated with mAb anti-SI antibodies. The samples were analysed by SDS-PAGE and phosphorimaging. (B) COS-1 cells were transfected with the cDNAs corresponding to the Ala mutants SIQ1098A and SIAla3Q to which YFP was fused (denoted pSIQ1098A-YFP and pSIAla3Q-YFP) and fixed 48 hours after transfection for immunofluorescence staining with calreticulin-, ERGIC 53- and GM130-specific antibodies. Alexa Fluor 633-conjugated antibody was used for immunodetection of the compartment-specific antibodies as indicated in Fig. 3. Arrows indicate YFP fluorescence on the plasma membrane. n, nucleus. Bars, 25 µm.





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