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. 2015 Oct 9;7(10):4054–4066. doi: 10.3390/toxins7104054

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© 2015 by the authors; licensee MDPI, Basel, Switzerland.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

The effects of Stx1, Stx2 and their A and B subunits on cytokine production in macrophage-like THP-1 cells. Macrophage-like THP-1 cells were incubated with 100 ng/mL of Stx1 or Stx2 or recombined Stx1 (Stx1R) or Stx2 (Stx2R) or were incubated with equimolar concentrations of their A or B subunits (3.125 nmol or 15.625 nmol, respectively). The recombined toxins were produced by incubating 5 mol of B with 1 mol of A subunit on ice overnight. Cell culture supernatant solutions were then collected and analyzed for cytokine/chemokine concentrations using Luminex multiplex analysis. The values represent the means of three independent trials performed in duplicate ± standard deviation for Stx1 and Stx2, or their A and B subunits. The assay was only performed once in duplicate for Stx1R and Stx2R.