Fig. 3.

Chemical cross-linking with DTME. Clone 22a cells transfected with 3 µg of FLAG–GIPC plasmids were incubated with 0.15 to 1.0 mg/ml of thiol-specific crosslinker DTME for 1 h at 4°C. Cells were lysed in PBS containing 1% Triton X-100 and analyzed by SDS–PAGE and immunoblotting with anti-GIPC antibody (A,B) anti-FLAG mAb M2 (C,D) under non-reducing and reducing conditions respectively. The arrow head indicates the immunoreactive band of approx. 115 kDa and of monomeric form of GIPC. The non-specific bands are indicated by the asterisks. The numbers on the right indicate the molecular mass (in kDa).