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. 2001 Jul 31;98(16):9098–9103. doi: 10.1073/pnas.151252898

Figure 2.

Figure 2

The SoHo domain is necessary and sufficient for flotillin binding. (A) Schematic diagram of FLAG-epitope-tagged expression constructs of CAP, CAP mutants, vinexinα, vinexinα mutants, and chimeras of CAP-vinexinα. (B) LacZ, FLAG-epitope-tagged CAP, CAPΔSoHo, CAPΔSH3 CAPSoHo, Vxα, VxΔSoHo and VxΔSH3 were overexpressed in HEK293T cells. Lysates were subject to SDS/PAGE and immunoblotted with monoclonal FLAG antibody to evaluate expression level (Left, arrowheads). Lysates were incubated with glutathione-agarose-bound GST-flotillin for 1 h at 4°C. Precipitates were subjected to SDS/PAGE and blotted with monoclonal FLAG antibody or with Anti-Cbl antibody (Right, arrowheads). (C) LacZ, FLAG-epitope-tagged CAP, Vxα, Vx-CAP, and CAP-Vx were overexpressed in HEK293T cells, and lysates were subject to SDS/PAGE and immunoblotted with monoclonal FLAG antibody to evaluate the expression level (Left). Lysates were incubated with glutathione-agarose-bound GST-flotillin for 1 h at 4°C. Precipitates were subjected to SDS/PAGE and blotted with monoclonal FLAG antibody or with anti-Cbl antibody (Right).