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. 2016 Mar 30;4:21. doi: 10.3389/fcell.2016.00021

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Copyright © 2016 Richter, Ventz, Harms, Mostertz and Hochgräfe.

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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(A) Western blot analysis of known downstream targets of TAK1. THP-1 monocytes were treated with 1 μM 5Z or vehicle for 1 h followed by stimulation with PMA for the indicated time periods. (B) Percentage of recovered colony forming units (CFU) of S. aureus infected THP-1 cells pretreated with 1 μM 5Z normalized to untreated THP-1 cells. Data is represented as mean ± S.D. (n = 3). (C) Count of THP-1 cells present in the cell culture supernatant in uninfected controls, following infection with S. aureus either pretreated with 1 μM 5Z or not, and uninfected controls pretreated with 1 μM 5Z. Data is represented as mean ± S.D. (n = 3). (D) Fluorescence distribution of PMA-differentiated adherent THP-1 cells infected with GFP-expressing S. aureus either pretreated with 1 μM 5Z or not and uninfected control cells as analyzed by flow cytometry. (E) ELISA-based quantification of secreted chemokines IL-8, MIP-1a, MIP-1b, and GROa following interaction with heat- inactivated S. aureus either pretreated with 1 μM 5Z or not compared to control THP-1 cells. (F) Light micrographs of THP-1 cells 3 days post PMA stimulation either with or without pretreatment with 1 μM 5Z. (G) Cell counts of THP-1 cells present either in the cell culture supernatant fraction (upper chart) or adhered fraction (lower chart) monitored over 72 h post PMA treatment in the absence and presence of different concentrations of 5Z. Data is represented as mean ± S.D. (n = 3). Statistical analysis of variance was performed by two-way ANOVA with Bonferroni post tests. ^*p < 0.05, ^**p < 0.01, ^****p < 0.0001.