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. Author manuscript; available in PMC: 2014 Sep 10.
Published in final edited form as: Nature. 2013 Nov 20;505(7481):103–107. doi: 10.1038/nature12729

Figure 4. Regulation of hRegIIIα pore formation.

Figure 4

(a-c) hRegIIIα pore formation is inhibited by lipopolysaccharide. (a) 10 μM hRegIIIα was added to liposomes composed of lipids from an E. coli total lipid extract or from PC/PS as a control. (b) 10 μM hRegIIIα was added to liposomes (100 μM lipid) in the presence of varying LPS concentrations. (c) 10 μM hRegIIIα was added to ∼104 cfu of log phase L. monocytogenes in the presence of varying LPS concentrations. The assay was carried out at 37°C for 2 hours, and surviving bacteria were quantified by dilution plating. Assays were done in triplicate. Results in a-care representative of two independent experiments.

(d-e) The hRegIIIα N-terminal prosegment limits toxicity toward mammalian cells. (d) hRegIIIα was added to MODE-K cells and cytotoxicity was determined by quantifying lactate dehydrogenase (LDH) release. LDH activity was assessed by spectrophotometric detection of an enzymatic product of LDH at 492 nm. (e) 10 μM hRegIIIα or pro-hRegIIIα was added to MODE-K cells and LDH release was quantified. Maximum LDH release was determined by treating cells with NP-40 detergent.