FIGURE 4.

Internalization of LGR5 is regulated by a motif between positions 834 and 869. Shown are primary amino acid sequences of the C-terminal tail for each construct (canonical GPCR NPXXY domain in gray). HEK 293T cells were transiently transfected with the indicated 3×HA N-terminally (red) and C-terminally EGFP (green)-tagged Lgr5 constructs: FL-LGR5 (A), 869 (B), 874 (C), or 902 (D). Cells were pulsed with a MαHA antibody for 45 min on ice, washed, chased for 0, 7.5, 15, 30, or 120 min at 37 °C, fixed, permeabilized, and stained with a GαM568 antibody (red). Merged 100× confocal images are presented (blue, nuclear counterstain).