Immunofluorescent staining of retinal microglial cells. (a) Microglial cells were transferred into a 16-well chamber slide and stimulated with LPS in the absence of L929 supernatant for 24 h. After stimulation, cells were stained for activation markers, including MHC-II (FITC), CD86 (PE), and CD40 (APC). The slides were observed by confocal microscopy. (b) Microglial cells were transferred into a 16-well chamber slide, incubated with POS-FITC at the ratio of cells to POS-FITC 1 : 5 for 18 h, and stained for CD11b (APC). (c) Microglial cells (1 × 104/well) were transferred into a 96-well plate and stimulated with 1 μg/mL of LPS for 24 h. The supernatant was collected and measured for the presence of IL-1β, IL-12, IL-10, CCL2, GM-CSF, IL-6, TNF-α, and CCL5 using CBA. *P ≤ 0.05 compared with control nonstimulated cell supernatant. Mean ± SEM, n = 3.
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