fig2

Figure 2. Microglia differentially express inflammation-associated genes in three neuroinflammatory models that demonstrate robust microglial reactivity. A: representative images of surveying microglia from the cortex of naïve mice and reactive microglia from LPS 24 h, 3 week Cuprizone, and EAE Early 3 & 4 mice; B-D: analysis of NanoString data of 248 differentially expressed inflammation-associated genes in CD11b⁺ cells. Background subtraction was performed using the maximum value across samples of the negative controls and data normalization was performed using the geometric mean expression of six internal reference genes (CLTC, GAPDH, Gusb, Hprt, Pgk1, Tubb5). Reporter probe counts reflecting the numbers of mRNA transcript in the RNA sample were analyzed and quantified using the nSolverTM Analysis Software, and are represented by fold-change compared to naïve cells. Two mice were pooled per sample and three total samples per group were submitted for NanoString analysis. Microglia were isolated by CD11b Miltenyi beads from the cortex of mice induced with EAE, Cuprizone, or LPS at early time points where neuronal pathology was detectable but had not peaked (EAE Early 3 & 4^[79], 3 week Cuprizone^[123], LPS 24 h)^[80]; B: heat map of differentially expressed genes; C: venn diagram representing the number of genes that are significantly upregulated, 1.3 fold-change or greater, in microglia from mice induced with EAE, Cuprizone, or LPS; D: venn diagram representing the number of genes that are significantly downregulated, 1.3 fold-change or greater, in microglia from mice induced with EAE, Cuprizone, or LPS; E: table showing the number of genes that were significantly upregulated (upward arrow) or downregulated (downward arrow) in each experimental group, and the number of altered genes shared among groups. Scale bar = 10 µm. P < 0.05. LPS: lipopolysaccharide; EAE: experimental autoimmune encephalomyelitis