DCIR internalization is clathrin-dependent. (A) For further analysis of DCIR endocytosis, a CHO cell line stably expressing human DCIR-EGFP was generated. DCIR expression was detected by FACS (top panel), either via EGFP detection (left, dotted line represents untransfected cells) or after labeling with anti-DCIR antibodies (right, dotted line represents isotype control). DCIR could also be detected by CLSM (bottom panel, left) and Western blot analysis (right). (B) DCIR internalization is inhibited by DN-dynamin. Cotransfection of the DCIR expressing CHO cell line with DN-dynamin constructs reduces DCIR endocytosis by 50% compared with cells cotransfected with WT-dynamin (*P ≤ .009) or to mock-transfected cells (**P ≤ .05). The data shown represent the mean (± SEM) of 3 independent experiments.