Fig. 1. Cellular localization of full-length MAN1 and CHL. (A) Proteins expressed by the plasmid constructs FL and FG-CHL, which were used to transfect COS-7 cells. Domains of proteins are represented as follows: yellow, FLAG-tag; white, nucleoplasmic, luminal and C-terminal domains of MAN1; red, LEM domain; orange, the first transmembrane segment of MAN1; dark green, the second transmembrane segment of MAN1; pink, transmembrane segment of CHL; light blue, luminal domain of CHL. The N-terminus of each protein is at the left. (B) Nuclear rim fluorescence characteristic of inner nuclear membrane labeling is seen for full-length MAN1 (FL) and a labeling pattern characteristic of outer nuclear membrane, ER and endosomes is seen for CHL (FG-CHL). Cells were fixed and immunostained with anti-FLAG monoclonal antibodies which were recognized by FITC-conjugated secondary antibodies. Bar, 10 µm.