HZ-induced ROS and IL-1β production is mediated by PKC, which is in turn target of <i>Leishmania</i> parasites induced dephosphorylation.

<p>(<b>A</b>) PMA-differentiated THP-1 cells (1x10⁶ cells/mL) were stimulated with 200 μg/mL of HZ for indicated time points. (<b>B</b>) THP-1 or LPS-primed BMDMs (0.1x10⁶ cells/100 μL) were pre-treated with indicated concentrations of PCK inhibitor GÖ6850 for 30 min. After 1.5 hrs, ROS production was measured by fluorescence using DCFA. Data shows mean values with SEM of 3 independent experiments. Differences were considered significant for p < 0.05. (#) denotes significant changes between uninfected and HZ treated samples. (*) denotes significant changes of ROS production due to PKC inhibition Cells were pre-infected with the indicated <i>Leishmania</i> species for 2 hrs (<b>D</b>, <b>E</b>) or pre-treated with the indicated amounts of the PCK inhibitor GÖ6850 for 30 min (<b>C</b>) and then stimulated or not with 200 μg/mL of HZ for 6 hrs (<b>C</b>) or as indicated (<b>D</b>). Cell extracts of PMA-differentiated THP-1 were pre-infected with <i>L</i>. <i>major</i> GP63 wild type (<i>Lmj WT</i>) or <i>L</i>. <i>major</i> GP63^-/- (<i>Lmj KO</i>) parasites for 2 hrs and then stimulated or not with 200 μg/mL of HZ for 1 hr (<b>E</b>) Cell extracts were collected and subjected to Western blot analysis with the indicated antibodies (<b>A</b>, <b>C</b>, <b>D</b> and <b>E</b>). (-) indicates untreated samples. (PKCsub) denotes the usage of an antibody specific for PKC-dependent phosphorylation.