Fig. 8

The activation state of OIR microglia is altered by Tregs. a Representative confocal image of a flat mounted retina from Foxp3^rfp mice at postnatal day (P) 18 with OIR and treated with the IL-2/anti-Il-2 mAb complex. Foxp3⁺ cells (red, arrowheads) contact microglia (Iba1 immunolabeling in green, arrows). Scale bar, 40 μm. b–e Microglia from retinas of OIR mice at P18 were co-cultured with Tregs purified from adult mouse spleen and incubated with a blocking anti-CTLA-4 mAb (10 μg/ml). Flow cytometry was performed and mean fluorescence intensity (MFI) quantitated for b CD40, c CD80, d CD86, and e CD11b. Fluorochromes are allophycocyanin (APC), PerCp-Cy5.5, phycoerythrin (PE), and AF700. *P < 0.05, **P < 0.01, and **P < 0.001 to OIR control. ^# P < 0.05, ^## P < 0.01, and ^### P < 0.001 to OIR + Tregs. Data are representative of two independent experiments each containing three replicates (one-way ANOVA with Student’s t test). Values are expressed as mean ± s.e.m