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A. B. Supplementary Data Figure 1 A, Binding of Alexa 546-CRP ( red ) to COS-7 cells expressing LOX-1, SR-A, CD36, SR-BI, and dectin-1. b -galactosidase (lacZ) was used as a negative control. The nuclei of the cells were counterstained with DAPI( blue ).
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A B Supplementary Data Figure 1 A, Binding of Alexa 546-CRP (red) to COS-7 cells expressing LOX-1, SR-A, CD36, SR-BI, and dectin-1. b-galactosidase (lacZ) was used as a negative control. The nuclei of the cells were counterstained with DAPI(blue). B, Quantitative analysis of Alexa 546-CRP binding to COS-7 cells expressing scavenger receptors and dectin-1.The binding was normalized with the expression level of the receptors. The asterisks represent a significant difference vs. negative control (*P<0.01). Method COS-7 cells transfected with the indicated cDNAs were washed with ice-cold DMEM/10 mM HEPES and chilled on ice for 30 min. Then, the medium was replaced with 3 mg/L Alexa 546-CRP-containing ice-cold DMEM/10 mM HEPES, and cells were incubated on ice for 1 h. After washing with ice-cold PBS, the cells were fixed with phosphate-buffered formalin (Wako) and permeabilized with 0.1% Triton X-100/PBS. The expression of each receptor was assessed by immunostaining with anti-V5 antibody (1:1000, Nacalai tesque) combined with Alexa 488-anti-mouse IgG (1:2000, Invitrogen).
