Fig. 5: PPARα and PPARα/γ-dual agonists enhance, whereas PPARγ agonist delay bacterial (AIEC-LF82) clearance.
a Schematic displays the experimental design and workflow. Thioglycolate-induced murine peritoneal macrophages (TG-PM) pretreated with PPAR agonists (see box, below; 20 nM GW7647, 10 μM Pioglitazone and 1 μM PAR5359) were infected with AIEC-LF82 (MOI 10) and subsequently analyzed for the bacterial count (Gentamicin protection assay), generation of cellular ROS, secretion of inflammatory cytokines (in supernatant media by ELISA) and the induction of cytokines (gene transcript analysis by qPCR, the cycle threshold (Ct) of target genes was normalized to 18S rRNA gene and the fold change in the mRNA expression was determined using the 2−ΔΔCt method). b Line graphs (left) display percent viable bacterial counts at indicated times after infection. Bar graphs (right) display the AUC. c Line graphs (left) and bar graphs (right) display the extent of ROS generation over time. d Bar graphs display the relative expression of transcripts of multiple cytokines (IL1β, IL6, TNFα, and IL10). All results are displayed as mean ± SEM. (n = 3). Significance: n.s, not significant, p-value < 0.05 was considered significant. e Line graphs (left) and bar graphs (right) showing the levels of indicated cytokines secreted in the media. Statistics: All results are from at least three independent experiments and results displayed as means ± SEM. Significance was tested using two-way/one-way ANOVA followed by Tukey’s test for multiple comparisons. All results are displayed as mean ± SEM. (n = 3). Significance: ns, non-significant, p-value < 0.05 was considered significant. See Supplementary Fig. 11 for similar bacterial clearance assays performed using Salmonella enterica.
