Figure 1.

Molecular cloning of LOX-1. (A) Ba/F3 cells were retrovirally transfected with a cDNA library from the variant Ba/F3 (#BEB1) cell line, immunologically stained for Ecrg4–Fc, sorted with a fluorescence-activated cell sorter, expanded in culture, and sorted again. The Ecrg4–Fc staining profiles of the Ba/F3 cells (1^st sort) and the cells after two cycles of sorting (3^rd sort) are shown. (B) The genes derived from the cDNA library were amplified with PCR using single-cell clones stained for Ecrg4–Fc from the P2 area of the 3^rd sorted cells. The amplified fragment (indicated by an arrow) was sequenced. LOX-1 cDNA, including the full-length coding region, was isolated from all three single clones. (C) Schematic representation of mouse LOX-1 and its truncated mutant. TM, transmembrane domain; CTLD, C-type lectin-like domain. (D) HEK293 T cells transfected with Flag-tagged wild-type mouse LOX-1 (left) or a CTLD-deletion mutant of LOX-1 (right) were incubated with Ecrg4–Fc before cell fixation. The binding of Ecrg4–Fc was detected with a FITC-labelled anti-human IgG Fc antibody (green; Fc), and the Flag-tagged LOX-1 expression was detected with an anti-Flag antibody (red; Flag). Nuclei were stained with Hoechst (blue). Scale bar 20 μm.