Fig. 6

miR-27b regulates inflammatory response in ischemic and tumor tissue. (a, b) IHC and IF for F4/80 of tissue sections harvested from HLI model (a) and LLC tumors treated as indicated. Cell nuclei are visualized with DAPI (blue). (c-d) quantification of the cells positive for macrophage marker, F4/80, (c) Macrophages were counted using ImageJ software (National Institutes of Health). Data is presented as box plot with whiskers and P values calculated using Mann–Whitney method. (d) Macrophages were counted using “object count” function of Nikon Elements software and statistical analysis performed using Wilcoxon test as above. (e-g) Cultured macrophages (RAW 264.7) were electroporated with negative control RNAi, miR-27b mimic (M) and inhibitor (I). Gene expression was measured by real-time PCR with L19 and beta macroglobulin as internal controls. PPARγ (e), a miR-27b target, and inflammatory chemokines IL-10 (f) and IL-12 (g) were assessed by real-time RT-PCR and normalized against untreated control (NT no treatment). The results of three independent experiments are pooled together. Statistical significance was assessed by Wilcoxon Signed Rank test, as above