Figure 8

cDN-induced repolarization of M2 polarized macrophages towards the M1 pro-inflammatory subtype. To establish M2 subtype of macrophages, cells were pre-treated with 40 ng/ml IL-4 for 48 hrs prior to the addition of cDNs. (a,b) A 6-hr treatment with either 2′3′-cGAMP or c-di-AMP significantly decreased Arg-1 and Fizz1 mRNA expression as compared to the IL-4-alone treated cells. (c–f) Moreover, IFN-β, CXCL-10, iNOS and IL-12p40 mRNAs were also increased at either the 6 hr or the 24 hr timepoint post-cDN exposure. All cDN treatments were carried out using lipofectamine permeabilization of the cultured cells. Significance was assessed by one-way ANOVA followed by Tukey’s multiple comparisons post-hoc test between all groups. Values represent the means ± SEM of the fold changes in the respective mRNAs; *P < 0.05, **P < 0.01, ***P < 0.001 vs. the control groups treated with IL-4 alone. Data was normalized to β-actin mRNA and experimental transcripts expressed as the relative fold change in each mRNA tested.