Fig. 8. Analysis of Triton solubility of GFP-22-Nglyc and GFP-22-MutNglyc. (A) Detergent lysates prepared from MDCK cells expressing GFP-22-Nglyc (upper part of panel) or GFP-22-MutNglyc (lower part of panel) were brought to 40% sucrose and analysed by flotation through a sucrose step gradient, as described in Materials and Methods. Half of each fraction was analysed by western blotting, using anti-GFP, anti caveolin I or anti-E cadherin antibodies, as indicated. (B) Half of fraction 4 and 1/20th of fraction 11 (11^*) from the same gradient of panel (A) were analysed by western blotting with anti-GFP antibodies. Bracket, arrow and arrowhead in (A) and (B) have the same meaning as in Fig. 7C. Note enrichment of mature glycosylated form of GFP-22-Nglyc in fraction 4, which corresponds to the interface between the 30% and 5% sucrose layers and contains raft-associated caveolin.