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. 2020 Mar 5;10(1):171–190. doi: 10.1016/j.jcmgh.2020.02.006

Figure 3.

Figure 3

Stx2a induces apoptosis and necrosis in isolated mesenchymal cells. (A) Stx2a induces apoptosis in isolated mesenchymal cells. Mesenchymal cells derived from mRuby2 E-cadherin HIOs challenged by addition of Stx2a (30 ng) were incubated for 27 hours. Cells were stained for apoptosis (Caspase 3). Scale bar: 100 μm. (B) Caspase 3 quantification. Fluorescence intensity was quantified using ImageJ software and plotted as means ± SD, n = 3. Statistical significance was assessed using a 2-tailed, unpaired t test. (C) Stx2a induces necrosis in isolated mesenchymal cells. Mesenchymal cells derived from mRuby2 E-cadherin HIOs were pretreated with or without 2 μmol/L antinecrotic factor Necrox-5 for 1 hour before addition of Stx2a (30 ng). The cells were incubated for 27 hours and stained for Sytox-green. Scale bar: 50 μm. Representative images of experiments performed in triplicate. (D) Sytox-green quantification. Fluorescence intensity was quantified using ImageJ software and plotted as means ± SD, n = 3. Statistical significance was assessed using a 2-tailed, unpaired t test.