Fig. 4

Caffeine ameliorates LPS-induced astrocyte activation in LRRK2 G2019S KI mice. (A) Mice of both genotypes were treated with a single dose of LPS (5 mg/kg, i.p.) or caffeine (daily; 20 mg/kg, i.p.). Body weights were monitored daily for two weeks. The percentages of initial body weights are shown. The number of mice was as follows: WT–LPS,  n= 4; LRRK2 G2019S–LPS, n = 6; WT–LPS + caffeine, n = 6; and LRRK2 G2019S KI–LPS + caffeine, n = 7. One WT mouse in the LPS-treated group died on day two post-LPS injection and was excluded from the study. (B) Representative blot of GFAP expression levels in striatal extracts are shown. (top). The corresponding quantifications using all biological replicates are shown below. n = 4–6 mice brain lysates per group. NS = normal saline group (C) Brain sections containing the substantia nigra were immunostained with an anti-GFAP antibody (green). Representative confocal images (top) and quantification of the GFAP⁺ area normalized to the total area are shown (bottom). Scale bar, 100 μm. (D-E) Striatal extracts were subjected to Western blot analysis to measure P-LRRK2 (Ser1292) (D) and P-Rab12 (T72) (E) expression levels. The corresponding quantifications are shown (below). n = 4–7 mice per group. For (B-E), *P < 0.05; **P < 0.01; ****P < 0.0001, n.s.: nonsignificant; one-way ANOVA followed by Tukey’s multiple comparison post hoc test. The data are presented as the means ± SEMs