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. 2020 Jul 7;11(4):e01470-20. doi: 10.1128/mBio.01470-20

FIG 11.

FIG 11

Shiga toxin production by lysogens. HIO mesenchymal cells were incubated for 18 h with LB broth or filter-sterilized supernatant from EHEC or Nissle::Stx2PT in the presence or absence of neutralizing antibodies to the Shiga toxin Stx2a and Stx2b subunits (30 ng each). (A to E) Cellular death was assessed by staining with Sytox green. Representative bright-field (BF) images with fluorescence are shown. Boxed regions represent the areas used to quantitate fluorescence. Bars, 50 μm. (A) LB control; (B) EHEC supernatant; (C) EHEC supernatant and anti-Stx2; (D) Nissle::Stx2PT supernatant; (E) Nissle::Stx2PT supernatant and anti-Stx2. (F) The fluorescence intensity in representative boxed regions was quantified using ImageJ software and plotted as the mean ± SD (n = 3). Statistical significance was assessed with GraphPad Prism (version 5) software, using one-way analysis of variance with Bonferroni’s posttest; comparisons between supernatant samples with and without antibody are indicated. **, P = 0.001 to 0.01.