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. 2018 Jul 31;7:135. doi: 10.1038/s41426-018-0135-9

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a rLep-OMP047-induced apoptosis of J774A.1 and THP-1 macrophages after exposure for the indicated times, determined by flow cytometry. The annexin V⁺/PI⁻ macrophages are early-apoptotic, while the annexin V⁺/PI⁺ macrophages are post-apoptotic/necrotic. rFliY, a recombinant expressed flagellum-associated cytosolic protein from L. interrogans, was used as the control. b Statistical summary of the early-apoptotic and post-apoptotic/necrotic percentages of rLep-OMP047-treated J774A.1 and THP-1 macrophages. Statistical data from experiments such as that shown in (a). Bars show the means ± SD of three independent experiments. *: p < 0.05 vs the early-apoptotic or post-apoptotic/necrotic percentages of rLep-OMP047-untreated macrophages. c Activation of caspase-8/-3 in J774A.1 and THP-1 macrophages treated with rLep-OMP047 for the indicated times, determined by fluorospectrophotometry. Bars show the means ± SD of three independent experiments. The fluorescence intensity (FI) values reflecting caspase-8 or caspase-3 activity in rLep-OMP047-untreated macrophages were set as 1.0. *: p < 0.05 vs the FI values reflecting caspase-8/-3 activity of rLep-OMP047-untreated macrophages. d Decrease in rLep-OMP047-induced J774A.1 and THP-1 macrophage apoptosis after Fas-antagonist-IgG/rLep-OMP047-IgG blockage, Fas depletion or caspase-8/-3 inhibition for the indicated times, determined by flow cytometry. Bars show the means ± SD of three independent experiments. *: p < 0.05 vs the early-apoptotic or post-apoptotic/necrotic percentages of rLep-OMP047-untreated macrophages (before treatment). ^#: p < 0.05 vs the early-apoptotic or post-apoptotic/necrotic percentages of Fas-antagonist-IgG- or rLep-OMP047-IgG-unblocked, Fas-undepleted and caspase-8/-3-uninhibited macrophages