Figure 9.

Microglial and macrophage cell depletion and autophagy after intracerebral injection of PGN. A) PGN or PBS was stereotaxically injected into the mouse CPU. At indicated time points after injections, coronal brain slices of 2.0 mm containing the CPU of one hemisphere were dissected. Tissue was homogenized, and CPU cells were costained with anti-CD45 (FITC), anti-CD11b (APC), AnV-PE (AnV), and 7-AAD and analyzed by flow cytometry gating on CD11b⁺ CD45⁺ microglial and macrophage cells. Bar graphs represents means ± sd of frequency of dead cells of 3 separate experiments. B) At indicated time points after injections, 10-μm brain sections were stained with an anti-CD45 (green) primary antibody and then with an Alexa Fluor 488 secondary antibody. Slides were analyzed under a laser scanning confocal fluorescence microscope. Arrowheads indicate parenchymal microglial and macrophage cells (CD45⁺ stained cells). Bar graphs represent means ± sd of numbers of CD45⁺ cells obtained from 10 fields/slide, analyzing the next 3 sections without the needle artifact of 3 separate experiments. *P < 0.05, ***P < 0.001 vs. PBS-injected mice.