Fig. 7

In vitro Tregs alter the activation of retinal microglia. a Primary cultures of microglia were established from the retinas of 10 to 12-day-old C57BL/6J mice. Quantitative real-time PCR revealed that ccl22 mRNA levels were increased by hypoxia (0.5% O₂). *P < 0.05. b–g Retinal microglia were exposed to hypoxia (0.5% O₂) and co-cultured with Tregs and a blocking anti-CTLA-4 mAb (10 μg/ml). Flow cytometry was performed and data are expressed as mean fluorescence intensity (MFI) 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 normoxia control. ^## P < 0.01 and ^### P < 0.001 to hypoxia control. ^† P < 0.05, ^†† P < 0.01, and ^††† P < 0.001 to hypoxia + Tregs. In the cell supernatant, an ELISA was used to measure f TNFα and g IL-6 levels in cell supernatant. *P < 0.05 and **P < 0.01 to normoxia control. ^# P < 0.05 and ^## P < 0.01 to hypoxia control. ^† P < 0.05 to hypoxia + 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