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Figure 4 | Journal of Inflammation

Figure 4

From: Activation of conventional protein kinase C (PKC) is critical in the generation of human neutrophil extracellular traps

Figure 4

Effect of PKC β on NET formation in response to PMA and DAG analogue OAG: Cells were plated as described and pre-treated with increasing concentrations of LY333531 (PKCβ inhibitor) before treatment with either 10 nM PMA or 20 μM 1-oleoyl-2-acetyl-sn-glycerol (OAG) for 4 h. A) Pre-Treatment with increasing concentrations of LY333531 decreases NET formation in response to 10 nM PMA. Data show mean +/− SEM for n=6 independent experiments for PMA stimulated cells and n=3 for OAG stimulated cells. *indicates p<0.05, ***p<0.001. B). Pre-treatment with increasing concentrations of LY333531 completely abrogated NET formation in response to 20 μM OAG. Data show mean +/− SEM for n=3 independent experiments. *** indicates p<0.001. C-J). Pre-treatment with 1 μM LY333531 completely abrogated NET formation in response to PMA and OAG, images representative of n=3 independent experiments (C,D Cells treated with 10 nM PMA. E,F cells pre-treated with 1 μM LY333531 for 30 min then 10 nM PMA. G,H cells treated with 20 μM OAG. I,G Cells pre-treated with 1 μM LY333531 for 30 min then 20 μM OAG. K,L Higher magnification images of effects of treatment with 100 nM LY333531 (L) prior to 10 nM PMA. Fluorescent and phase images combined. There is an absence of spreading NET material in the LY333531 treated cells although some diffuse NET formation is still present at this concentration of inhibitor. Scale bars = 10 microns.

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