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Cbl-N binding site is mapped to a hydrophobic region in SLAP C terminus. COS-7 cells were transiently transfected with Cbl-N/pEBM and various SLAP wild-type or mutant constructs in the pEBG vector. Cell lysates were precipitated with glutathione-Sepharose. Protein complexes were separated by 10% SDS/PAGE and blotted with anti-GST and anti-myc antibody. The positions for GST-SLAP and myc-Cbl-N are indicated. Refer to Fig. 7A for abbreviations of SLAP mutants.
