Figure 4.

Gangliosides increase the detergent solubility of GH-DAF. (A) MDCK GH-DAF cells were loaded with 100 μM bbG for 1 h at 37°C, with 100 μM NBD-C₆-HPC for 1 h at 8°C, with 100 μM GM₁ for 1 h at 37°C, or with 100 μM bbG for 1 h followed by a 6-h incubation period with serum containing DMEM (A) or treated with 10 mM methyl-β-cyclodextrin for 1 h at 37°C or incubated with anti-GH for 1 h at 12°C followed by incubation with secondary antibody (B). Cells were extracted with Triton X-114 for 30 min at 4°C and subjected to centrifugation, and GH-DAF in the soluble (S) and insoluble fraction (I) was detected as described in MATERIALS AND METHODS.