(A). HEK293 cells were transiently transfected with GLUT12-HA, GLUT12-AA-HA, or GLUT12-TQP-HA. Cells were stained with or without prior permeabilization using an HA-antibody. Decreased levels of plasma membrane staining were observed in GLUT12-AA or GLUT12-TQP-expressing cells compared to the cell surface expression levels of wild-type GLUT12. (B) Relative amounts of cell surface-associated GLUT12 were quantified using flow cytometry with HEK293 cells transiently expressing GLUT12-HA, GLUT12-AA-HA, or GLUT12-TQP-HA. In agreement with the immunofluorescence results, GLUT12-AA-HA and GLUT12-TQP-HA exhibit less cell surface expression than wild-type GLUT12. (C) GLUT12-HA and GLUT12-TQP-HA localized to similar compartments in endometrial stromal cells which endogenously express GLUT12. Images were taken at 60× magnification.