Fig. 4.

Agathisflavone modulates microglia-oligodendrocyte interactions. Organotypic cerebellar slices from P10-12 Sox10-EGFP mice were maintained for 7 DIV and then treated with LPC for 15–17 h, followed by agathisflavone (FAB) at 5 or 10 μM for a further 2 DIV, or 0.1% DMSO vehicle. (A) Photomicrographs of IBA1 immunostaining (red) and SOX10-EGFP+ oligodendrocytes (green) showing oligodendrocytes-microglia contacts in the different treatment groups; scale bar 20 μm. (B) Diagram illustrating microglial processes contacting oligodendrocytes body (Pr-B), or apposition of microglial and oligodendrocyte cell bodies (B—B). (C) Grouped bar graph showing the number of microglial contacts per SOX10+ cells; data are expressed as the mean ± SEM (n = 6), *p < 0.05, **p < 0.01, ***p < 0.001 (comparing control to treatment groups); ‡‡p < 0.01, ‡‡‡p < 0.001 (comparing LPC-DMSO to LPC+FAB5 and LPC+FAB10; two-way ANOVA followed by Tukey’s post-hoc test. (D) Photomicrographs of slices illustrating Sox10-RGFP + oligodendrocytes (green) and immunolabelling for MBP (red) and Iba1 (yellow), showing interrelationships between microglia, oligodendrocytes, and myelinated fibres in the different treatment groups; clusters of IBA1+ microglia around myelin debris and oligodendrocytes are evident following LPCv treatment and are rarely observed in controls or following agathisflavone treatment. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)