Fig. 5
Expression of a constitutively active form of IRF3 in Cx3cr1+ cells in the brain is sufficient to induce neuroinflammation. A Representative images of microglia (Tmem119+ cells were picked) co-stained with Iba1 and Sholl analysis performed using filament tracer software from Imaris. The scale bar is 21 μM. B Total volume of the cells did not change between any of the groups tested. C Quantification of the microglia morphology shows that microglia (Cells positive for Tmem119) from IRF3-2D,Cre_Tam group show significantly reduced branching compared to IRF3-2D,Cre_Oil and Cre_only groups. D Microglia from IRF3-2D,Cre_Tam group show reduced number of intersections at 10–30 μm compared to that of IRF3-2D,Cre_Oil and Cre_only groups. * represents comparison with Cre_only, # represent comparison with IRF3-2D,Cre_Oil. For Sholl analysis N=6-8 for each group. >7 microglia were analyzed per animal. Data was analyzed using One-way ANOVA. *p<0.05, **p<0.01, ***p<0.001, ****p<0.000. E-G Representative images and quantification of FACS analysis showing the presence of infiltrating myeloid cells (CD11b+,CD45high) in the IRF3-2D,Cre_Tam (Yellow arrow) group in addition to the microglia population (CD11b+,CD45intermediate) (White arrow). F Shows the quantification as % of the parent population, while G shows absolute number of infiltrating cells. N= 8,9 each group. One-way Anova with Sidak’s multiple comparison test. ****p<0.0001.H-I Quantification of CD45 and CD11b MFI gated on microglia (White arrow in E) shows significant upregulation in IRF3-2D,Cre_Tam, suggesting more reactive state compared to Cre_only controls. N= 8,9 each group. One-way Anova with Sidak’s multiple comparison test. ****p<0.0001. J-K Quantification of the western blots shows proliferation of astrocytes, as measured by GFAP levels in cortical lysates, of IRF3-2D,Cre_Tam compared to Cre_only. N= 6,7 each group. Kruskal-Wallis test with Dunn’s multiple comparison test. **p<0.01