Fig. 5. Treatment with IL-10⁺ B cells promotes M2 phenotype induction in microglia, post-MCAO.

Primary MG, isolated and cultured from MCAO-treated WT male and female mice, were harvested after 21 days in vitro (at confluency) and cultured in GM-CSF-free medium for 5 days. MG were stimulated with 10ng/ml LPS for 4 hours. Supernatants were discarded after 4 hours and one of the following treatments was given in 1mL of fresh culture medium: no treatment, 20ng recombinant IL-10 (rIL-10), 20ng/ml rIL-4 or IL-10⁺ B cells at a 1:1 ratio with MG. The MG cells were incubated with mentioned treatments at 37°C and 5% CO₂ for 24 hours and the M2 marker, CD0206 was determined by flow cytometry. Values are given as mean ± S.E.M. Data presented are representative of n = 2 separate co-culture set-ups, with each treatment condition done in duplicate for every experimental set-up. Statistical analysis was performed using One way ANOVA with post-hoc Dunnett test. Significant differences between sample means are indicated as *p≤0.05 and ***p≤0.001 as compared to the LPS-stimulated condition. Statistical differences between the two sexes were performed by two-way ANOVA followed by the post-hoc Bonferroni multiple comparison test, with #p≤0.05 as compared to the respective treatment in male MG