Figure 2

Induction of chemoatractants by LPS injection in the cerebellum. (a) The expression levels of proinflammatory chemokines, such as MCP-1 and MIP-1α, in the cerebellum at 1 day and 7 days post-LPS injection. MCP-1: ^**p = 0.003 versus CON (n = 3 per group, t(4) =  − 6.26). MIP-1α: ^**p = 0.008 versus CON, ^#p = 0.049 versus PBS (1 day) (n = 3 per group, F(4,10) = 6.042). (b and c) The expression pattern between glial cells (microglia and astrocytes; green) and proinflammatory chemokines (MCP-1 and MIP-1α; red) in the cerebellum at 1 day and 7 days post-LPS injection (Scale bar = 50 μm). MCP-1 and MIP-1α expression were upregulated by microglia and astrocyte in the LPS-exposed cerebellum (b), but rarely colocalized with astrocytes and microglia, respectively (c, arrowheads). Scale bar = 50 μm. (d and e) Increases in mRNA expression levels of MCP-1 and MIP-1α by treatment with LPS and LPS + IFN-γ in primary microglia (d) and astrocyte cultures (e). Relative mRNA expression of MCP-1 and MIP-1α at 6 h after treatment with inflammatory stimuli was determined by RT-PCR. Graphs represent quantitative analysis of gel images normalized to GADPH. Microglia: ^**p = 0.004 (MCP-1, t(4) =  − 6.003) and ^**p = 0.003 (MIP-1α, t(4) =  − 6.750) versus CON (n = 3 per group). Astrocytes: ^***p < 0.001 (MCP-1, t(4) =  − 74.306; MIP-1α, t(4) = − 22.787) versus CON (n = 3 per group). All values are expressed as the mean ± standard deviation (SD). Statistical significance tested with one-way ANOVA with Tukey’s post-hoc analysis (a; MIP-1α) and two-sided paired t-test (a; MCP-1, d, and e). The membranes were initially cut according to the molecular weight size of proteins. The images of blots were displayed in cropped format, and original blots of (a), (d) and (e) with cropped demarcation of membranes are shown in Supplementary Fig. 2.