Fig. 2

Dietary treatment with CSF1R inhibitor depletes IBA-1⁺ microglia from the ADR-treated hippocampus. Quantification for the IBA-1 immunoreactivity using immunofluorescence staining, laser scanning confocal microscopy and 3D volumetric-based analysis demonstrates that treatment with PLX5622 for 6 weeks eliminates IBA-1⁺ microglia from the ADR-treated (ADR + PLX5622) brains (green, IBA-1; blue, DAPI nuclear counter stain). a-c Representative confocal micrographs for IBA-1 immunostaining from the hippocampal dentate gyrus showing dentate hilus (DH), granule cell layer (GCL) and CA1 sub-regions from the control and ADR groups receiving control diet and, ADR-treated mice receiving PLX5622 diet (insert, high resolution image of IBA-1⁺ cell). d 3D algorithm-based quantification (Autoquant and Imaris) of IBA-1⁺ microglia show > 96% depletion in the ADR + PLX5622 group. Data are presented as mean ± SEM (N = 4–6 mice per group). P values are derived from ANOVA and Bonferroni’s post hoc test. *, ⁺ P < 0.0001 compared with Control and ADR groups respectively. Scale bars, 200 μm, a-c 5 μm, insert