Fig. 3. Scriptaid increases CD86 and iNOS and decreases GPR18, CD38 and FPR2 gene expression, whereas RGFP966 only increases CD86 gene expression. (A) Experimental set-up of the in vitro experiments: BMDMs were first pre-stimulated for 1 hour either with LPS (200 ng/ml; M1 macrophages) or with IL-4 (33 ng/ml) or IL-13 (33 ng/ml; M2 macrophages). Thereafter, the BMDMs were stimulated for 24 hours with scriptaid (0.2 µM and 1 µM) or RGFP966 (5 µM and 10 µM). (B~H) M1 macrophages treated with scriptaid or RGFP966 were lysed and RNA was collected for gene expression analysis of several M1 genes: CD86 (B), iNOS (C), IL-6 (D), IL-1β (E), GPR18 (F), CD38 (G) and FPR2 (H). CD86 and iNOS were increased after treatment with 10 µM scriptaid. However, GPR18, CD38 and FPR2 were decreased after treatment with 10 µM scriptaid. CD86 showed a slight increase in gene expression after treatment with 5 µM RGFP966, all other genes showed no significant difference after treatment with RGFP966. Data is shown as fold-change relative to ‘control+LPS’±SEM; *p<0.05; n=3 biological replicates. BMDMs, Bone marrow derived macrophages; LPS, lipopolysaccharide; IL-4, interleukin-4; IL-13, interleukin-13; CD38, cluster of differentiation 38; CD86, cluster of differentiation 86; FPR2, formyl peptide receptor 2; GPR 18, G protein-coupled receptor 18; IL-6, interleukin 6; iNOS, inducible nitric oxide synthase.
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