Fig. 4

Aldara treatment induces the infiltration of monocytes, but not neutrophils, into the brain. Mice were treated with 80-mg Aldara cream or control cream every 24 h for one, three or five consecutive days. Mice were euthanised 24 h following the final application. a Protein expression of CCL2 in whole brain lysate was assessed in control and treated brains 4 h, 12 h, 1 day, 3 days and 5 days after treatment. b Perfused brains were homogenised, and a single cell suspension was generated as described. Cells were labelled with fluorescent antibodies. CD45^hiCD11b⁺ cells were selected following dead cell exclusion and doublet exclusion, and representative gating strategy based on expression of Ly6C, Ly6G and CD64 is shown. c Counts per million cells were generated using total cell counts and are shown for neutrophils, monocytes, differentiating monocytes (population 2) and macrophages (population 3) at 1, 3 and 5 days following Aldara treatment. n = 4 mice per group. Significance was determined using one- (a) or two-way (b, c) ANOVA with Bonferroni multiple comparison post-tests. Data presented in a were log transformed prior to statistical analysis ****p < 0.0001 ***p ≤ 0.001 **p ≤ 0.01 *p ≤ 0.05