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. 2015 Sep 2;6:921. doi: 10.3389/fmicb.2015.00921

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Copyright © 2015 Popova, Teunis, Vaseghi, Zhou, Espina, Liotta and Popov.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Effect of Sterne and ΔNOS sups grown in microaerobic conditions on phosphorylation of AKT (S473) in HSAECs. The cells were exposed for 2 h to the sups of bacterial cultures grown for 24 h in static conditions. Corresponding pH values of the sups are shown. The lysates of the cells were used for western blots with a primary rabbit antibody against phosphorylated form of AKT (S473) and a secondary anti-rabit antibody conjugated with horseradish peroxidase. The activity of peroxidase in the bands was detected using a SuperSignal West Femto chemiluminescent substrate (Pierce) and quantitated with a digital camera. To take into account the effect of pH the ΔNOS sup after cultivation was titrated from pH 6.4 to pH 5.1 indicated by an asterik. Error bars show 95% confidence intervals obtained from three independent western blots. The preparation of the lysates was repeated twice with similar results.