Figure 4.

Immunoblotting of conditioned growth medium (A) and cell lysates (B) from CHO-K1 cells transiently transfected with wt or Gly1112Glu CUB 5–8 cubilin. Proteins were separated using SDS-PAGE (3–8%). and transferred to an Immobilon™ –FL PVDF transfer membrane using the iBlot™ Dry Blotting System. CUB 5–8 proteins were detected with a rabbit anti human cubilin antibody and visualised using LICOR IRDye λ 800 goat a-rabbit. Immunoreactive bands, consistent with the size of recombinant cubilin CUB 5–8, are indicated with white asterisks. No mutant CUB 5–8 protein was detected in the conditioned growth medium in contrast to the wt CUB 5–8 protein as previously described [39]. Both the wt and mutant protein were detected in cell lysates.