Figure 3 | Scientific Reports

Figure 3

From: The lipid components of high-density lipoproteins (HDL) are essential for the binding and transportation of antimicrobial peptides in human serum

Figure 3

Roles of lipids in the HDL for AMP-binding. (a) Purification of human serum components responsible for AMP-binding. The human serum components were pulled down by immobilized LL37, stepwise eluted by 30% and 70% TFE after overnight incubation at 4 °C, and analyzed by 15% non-reducing SDS-PAGE. (b) Lipid contents of the eluates (E1–E6). Significance is calculated according to One-way ANOVA test. *** indicates P ≤ 0.0005 which E5 compared with each other groups. (c) Effect of HDL, eluate E5 (top panel) and various lipids (bottom panel) on the antimicrobial activities of AMPs against E. coli (106 cfu/ml). Values are the mean ± SD (n = 3). Kruskal–Wallis tests were performed to determine the significance of the difference in each group. * Indicates P ≤ 0.05, ** Indicates P ≤ 0.005, *** Indicates P ≤ 0.0005, **** Indicates P ≤ 0.00005. (d) Band shift of AMPs by 10 µg HDL, E5 and various lipids (4 µg each, dissolved in 20 µg/10 µl BSA) and analyzed by 8% horizontal native PAGE and Coomassie blue staining. Arrow indicates control (non-shift) AMP. PC: phosphatidylcholine; SM: sphingomyelin; CO: cholesteryl oleate; BSA: bovine serum albumin.

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