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. Author manuscript; available in PMC: 2013 Aug 1.
Published in final edited form as: Microbes Infect. 2012 Mar 14;14(9):719–729. doi: 10.1016/j.micinf.2012.02.009

Figure 5. Analysis of Mycolactone – mediated cytopathicity on the fish macrophages.

Figure 5

[A] Cytopathicity on CLC cells, showing ethanol control treated cells [first left]; mycolactone AB treated cells [top panel] and mycolactone F treated cells [bottom panel]. The final concentration of mycolactone added to treated cells was 10μg. Scale bar = 50 μm [B and C] Cytotoxicity measured by LDH release and nucleosome enrichment. Culture supernatants were collected from wells containing CLC cells 24 h after treatment with mycolactone. [B] The amount of LDH released by necrosis was measured using a Cytotox 96 assay kit [Promega]. Data are means and standard deviations of the values obtained from triplicate samples; P=0.7, 0.03, 0.06 and 0.04 for 10ng, 100ng, 1μg and 10μg respectively [Student’s t test]. [C] Apoptosis was assessed at 24 h with the cell death detection enzyme-linked immunosorbent assay kit [Roche] and expressed as fold enrichment of nucleosomes. Data are means and standard deviations of the values obtained from triplicate samples; P=0.06, 0.9, 0.02 and 0.06 for 10ng, 100ng, 1μg and 10μg respectively [Student’s t test].