Fig. 2

Gene expression profile of CD163⁺ macrophages after brain ischemia. a) We isolated CD11b⁺CD163⁺ BAMs and CD11b⁺ CD163⁻ microglia by FACS from control rat brain and obtained RNA for gene expression analysis. Colors for cells in the drawings are arbitrary. b) By qRT-PCR we validated that sorted macrophages, but not sorted microglial cells, express Cd163. The expression of Aif1 (Iba-1) is higher in microglia than macrophages, whereas the expression of Siglec1 (CD169) is lower in microglia. Values are expressed as fold versus the mean value of CD163⁺ macrophages and are the mean±SD of n = 3–6 samples per group. **p < 0.01, *p < 0.05, two-tailed Mann-Whitney test. c) RNA was extracted from CD163⁺ cells immunosorted from the control and the ischemic rat brain at 16 h of reperfusion (n = 3 per group) to study ischemia-induced changes in gene expression profile using Affymetrix microarrays. The global heat map, where each lane represents macropahge gene expression from the brain of different control or ischemic rats, shows results of the microarray analysis with∣logFC∣> 2 and FDR < 0.01. d) Top diseases/functions associated to gene expression profile changes were obtained with ingenuity pathway analysis (IPA) gene ontology algorithms showing category scores. The minimum significance level (−log(p-value)) from Fisher’s exact test is indicated by the threshold (arrow). Inflammatory and immune functions are highlighted suggesting the involvement of CD163⁺ macrophages in such responses after brain ischemia