Figure 4
From: Agricultural dust derived bacterial extracellular vesicle mediated inflammation is attenuated by DHA
Stimulatory effects of EVs are sensitive to heating but are not due to LPS contamination and the effects of complete DE are partially due to the EV cargo. Saturated (100%) DE was subjected to heat (90 °C for 15 min) or LPS ablation (polymyxin B, 12 h @ 4 °C). HBEC were challenged with EVs isolated from these preparations (5% DE equivalents) or with heat-or polymyxin B-treated 5% DE for 24 h (A). Polymyxin B treatment significantly depleted LPS from EV suspensions (B). Alternatively, cells were challenged with complete 5% DE, an equivalent concentration of DE-derived EVs, or DE devoid of EVs, for 24 h. Supernatant medium was assessed for IL-6, IL-8 and AREG levels by ELISA (C). EV abundance (NTA) as a proportion of total protein for 5 different preps is shown in (D). Data shown for (A) and (C) are means of 3 independent experiments (8 or 14 technical replicates per condition). *p < 0.05, **p < 0.01, ***p < 0.0001 vs control (AVOVA, Tukey’s post-hoc test). For (B) and (D), 6 technical replicates per condition, ***p < 0.001, Student’s t test.
