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Figure 1


Fig. 1. OxLDL binding to HeLa cells expressing LOX-1. (A) Domain structure of the human LOX-1-FLAG construct. CD, cytoplasmic domain; TMD, transmembrane domain. (B) Equal quantities of total cell lysate (30 µg) from mock-transfected and LOX-1-FLAG transfected cells were analysed by western blotting using sheep anti-LOX-1 antibodies or sheep antibodies that recognise a ubiquitous Golgi membrane glycoprotein and loading control (TGN46). (C) Mock and LOX-1-FLAG transfected HeLa cells were incubated with labelled DiI-OxLDL ligand (red) prior to cell fixation, processed and stained with a mouse anti-FLAG antibody. Ligand binding was also performed after pre-incubation with the JTX92 LOX-1 blocking antibody. Cell surface LOX-1 was detected by the mouse anti-FLAG and FITC-conjugated anti-mouse IgG antibodies (green). (D) HeLa cells transfected with LOX-1-FLAG were incubated with DiI-OxLDL (red) in DMEM prior to washing and chasing for different time periods before fixation. Nuclei were stained with DAPI (blue). Scale bar: 20 µm.





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