Fig. 6
Phagocytic activity of microglia from Socs3fl/fl and DKO mice. (A) Microglia from Socs3fl/fl or DKO mice were seeded into 96-well plates. 24 h later, E.coli Alexa 594 Bioparticles or pHrodo E.coli Bioparticles were added into the wells. Fluorescence intensities were measured by plate reader and images were taken by fluorescence microscopy at the end of the study. (B) Fluorescence conjugated Alexa Fluor 594 E.coli particles were added to microglia for 30 min and the fluorescence intensities (B-1) were measured using Fluostar Omega microplate reader. After that, images were taken by Dmi8 fluorescence microscopy (B-2). (C) The pH sensitive E.coli particles were added to microglia. Fluorescent intensities (C-1) were measured using the Fluostar Omega microplate reader 0.5 h, 1 h, 2 h, and 3 h later. At the end of the study, images (C-2) were taken using Dmi8. Mean ± SD, N = 5 (B) and 3 (C). * P < 0.05, ** P < 0.01. Independent sample t test (B); Two-way ANOVA followed by Turkey’s test (C). Scale bar: 10 μm. (D) A confocal image from an aged DKO eye section showing cone arrestin (red) and GFP+ subretinal microglia (green). (D-1) Zoomed view of the marked area in (D) showing cone arrestin antigens (white arrows) inside a GFP+ filtrating cell. IS – inner segments of photoreceptors; ONL – outer nuclear layer