Fig. 3

Glycine-rich domain (GRD) in GAREM2 is necessary for its unique subcellular localization dependent on EGF stimulation. a Amino acid sequences of the N-terminal region of GAREM1 (upper) and GAREM2 (lower) are indicated as numbers. GAREM2-specific glycine-rich domain (GRD) is indicated by broken line. b Schematics of the primary structure of full length GAREM2 (upper) and its deletion constructs lacking GRD (∆GRD186-209 and ∆GRD172-209). c All recombinant proteins were expressed as the N-terminal GFP-fused form. Each construct was transfected into COS-7 cells. Confirmation of expressing GAREM proteins by immunoblotting using with GFP antibody. d COS-7 cells expressing GFP-GAREM2 proteins with [EGF( +)] or without [EGF(−)] EGF stimulation for 30 min were immunofluorescently stained using anti-EGF receptor antibody. Subcellular localization of the full-length and deletion mutant constructs of GFP-GAREM2 (green) and EGFR (red). Representative results are shown. Merged images are indicated in the right panels. Scale bars = 10 μm. e Extent of aggregation of wild-type and mutant GFP-GAREM2. Approximately 100 GFP positive cells were observed. Data are means ± S.E. (n = 5). *P < 0.05