Fig. 2

IKK2-CA animals exhibit prominent cerebellar neuroinflammation including microgliosis and astrogliosis. a-c Activation/infiltration of CD45 positive immune cells at different ages (8–36 weeks) in the cerebellum of IKK2-CA animals. d Staining for the myeloid cell marker CD11b indicates massive microgliosis at 12 weeks of age. Higher magnification (right panels): microglia display arborized morphology; highly activated microglia/infiltrated macrophages are rounded (amoeboid) cells. e Infiltration of Th-cells, shown by CD4 staining (age 12 weeks); right panels: higher magnification. f-i IKK2-CA induces a proinflammatory gene expression profile in the cerebellum of IKK2-CA mice increasing with age (qRT-PCR). Expression of chemokines (f), cell adhesion molecules (g), inflammatory cytokines (h) and the acute phase response factor Lcn2 (i) is elevated in IKK2-CA mice. Expression levels presented relative to HPRT (mean +/- s.e.m.); statistical analysis: 2-tailed Mann-Whitney-test (n = 4–7), ns: not significant (p > 0.05); * p < 0.05; ** p < 0.01. j Immunoblotting for Mac2 and GFAP indicates microgliosis and astrogliosis respectively, transgene expression (IKK1/2 immunoreactivity) and induction of the NF-κB target Lcn2. Representative immunoblot and quantification of GFAP immunoreactivity normalized to ERK2 (loading control), shown as mean +/- s.e.m. relative to Co 10w, statistical analysis: 2-tailed unpaired t-test (n = 3–4), ns: not significant; * p < 0.05; ** p < 0.01; *** p < 0.001. Images (a-e) show DAPI co-staining. Scale bars: (a-c) 100 μm; (d-e) 100 μm, right panels 25 μm