KLRL1 is expressed as a membrane-associated glycoprotein. (A) hKLRL1 transiently expressed in 293T cells is an N-linked glycoprotein. Lysates of 293T cells transiently transfected with phKLRL1/Flag or mock vector were analyzed by Western blot under nonreducing (Non-R, in the absence of 2-mercaptoethanol), reducing (Red, in the presence of 2-mercaptoethanol), or N-glycosidase F digestion (N-Gly) conditions. (B) Both human and mouse KLRL1 is expressed as a transmembrane protein. 293T cells and L929 cells were transiently transfected with phKLRL1-GFP (i), pmKLRL1-GFP expression vectors (iii), or GFP-alone control vectors (ii,iv), respectively, and fluorescence confocal microscopy analysis was performed 48 hours after transfection. Original magnification, ×900.