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Figure 4


Fig. 4. Porcupine promotes the lipid-modification of both wild-type Wnt1/Wnt3a and mutant Wnt1/Wnt3a, which possess C-to-S point mutations in the cysteine that is known to be palmitoylated. 293T cells were transfected with Wnt1, Wnt3a, Wnt1(C93S) or Wnt3a(C77S) in the presence of GFP (control) or mPorcD. Cells were lysed 24 hours post-transfection and extracted with TX-114 before analysis by SDS-PAGE and western blot. The blots were probed with Wnt1 (left), Wnt3a (right) and tubulin antibodies. Monomeric Wnt1/Wnt3a proteins migrate at ~40 kDa, whereas tubulin migrates at 50 kDa. In the absence of ectopic porcupine, Wnt1 and Wnt3a are found in both aqueous (A) and detergent (T) phases. The presence of ectopic porcupine promotes the partitioning of Wnt1, Wnt3a, Wnt1(C93S) and Wnt3a(C77S) into the hydrophobic phase.





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