Skip to main content
. 2021 Feb 18;12:633696. doi: 10.3389/fmicb.2021.633696

FIGURE 4.

FIGURE 4

A. fumigatus-induced EETs release requires PI3 kinase activation. Human eosinophils were pretreated with (A,B) the pan-PI3K inhibitor wortmannin (wortm – 100 ηM), (C,D) a class I PI3K inhibitor (AS605240 – 10 μM) and a PI3Kδ inhibitor (IC87114 – 1 μM), or their vehicle (DMSO), for 30 min and were subsequently stimulated with A. fumigatus conidia (cell: fungus ratio of 1:10) for 6 h. (A,C) EETs were quantified in the samples using a fluorescence microplate reader after Sytox Green addition (5 μM). The graphs represents the mean ± SEM of 3 (wortm), 3 (AS605240) and 7 (IC87114) independent experiments from different donors. *P < 0.05 and **P < 0.01 compared to the non-stimulated sample (medium); # P < 0.05, ##P < 0.01, ### P < 0.001 compared to AF + vehicle condition; one-way ANOVA followed by the Newman-Keuls test. (B,D) Confocal fluorescence microscopy of human eosinophils pretreated with (B) 100 ηM wortmannin, (D) 10 μM AS605240 and 1 μM IC87114 or their respective vehicle dilutions (DMSO) for 30 min and subsequently stimulated with A. fumigatus conidia (cell: fungus ratio of 1:10) for 6 h after staining for DNA (Sytox Green, green). Representative images of 3 (wortm), 3 (AS605240) and 7 (IC87114) independent experiments (n = 3, 3 and 7). AF = Aspergillus fumigatus.