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Fig. 2. DCC is palmitoylated. (A) Schematic representation of DCC. (B) Wild-type DCC, mutant C1121V DCC, or Fyn-transfected HEK293 cells were labelled with [³H]palmitate. Cell lysates were immunoprecipitated as described in the Materials and Methods and analysed by both autoradiography and western blotting. (C,D) HEK293 cells were transiently transfected with wild-type DCC or the DCC C1121V mutant. (C) Rafts were then isolated as in Fig. 1A and analysed for DCC, Fyn, Rab5 and TfR expression by western blotting. (D) Raft-containing fractions (Raft) and heavy fractions (HF) were pooled as in Fig. 1C. A quantification of DCC in raft fractions was performed using the GeneTools software. Right: a flow cytometric analysis of DCC expression performed on non-permeabilized wild-type DCC- or DCC C1121V mutant-transfected HEK293 cells (Immunostaining performed using anti-DCC antibody). The experiments shown in C and D are representative of at least three experiments.

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