Figure 5. Delineation of endocytic sorting motif in Kremen1.

HeLa cells were transfected with (A) Tac-Kremen1 (PLV-AAA), (B) Tac-Kremen1 (PLV-PAA), (C) Tac-Kremen1 (PLV-AAV), (D) Tac-Kremen1(PLV-ALA), or (E) Tac-Kremen1 (PLV-PLL). Transfected cells were starved in DMEM, 0.1% BSA, and 10 mM Hepes (pH 7.5) at 37°C. After washing with cold PBS, cells were incubated with mouse monoclonal anti-Tac and transferrin-Alexa488. Cells were chased at 37°C for 15 minutes. Cells were fixed, permeabilized, and stained with anti-mouse-IgG-Cy3. Scale bar 10 µm.