Figure 2

Endothelial association and transfection activities of three LNP formulations with different cationic lipid compositions. (A) Specificity and extent of cell association of LNP to HUVEC, as determined by flow cytometry. TNF-α-activated HUVEC were incubated with uLN or AbLN of three formulations for 3 h. The specificity of cell association to VCAM-1 was investigated by co-incubation with an excess of anti-VCAM-1 antibodies together with AbLN. Data are presented as normalized mean fluorescence intensity (MFI) ± SD values of three independent experiments. HUVEC_resting/HUVEC_{activated only} = 1. (B) Transfection activities of uncoupled and antibody-coupled LNP formulations loaded with siRNA_RelA in activated EC. TNF-α-activated HUVEC were transfected with uLN or AbLN containing siRNA_RelA at 100 nM concentration for 48 h and next processed for qPCR as described in the Section 2. Data are shown as relative fold change in gene expression ± SD of 3 independent experiments.