Fig. 4

JQ1 treatment inhibits microglial expression of inflammatory cytokines in the rd10 retina. Intravitreal injection of JQ1 (or vehicle) was performed at PN14, as described in Fig. 1. Retinas were collected at PN24. a Homogenates were prepared from retinas (3 groups, total 12 mice) and used for qRT-PCR. Quantification: normalization to GAPDH mRNA; mean ± SEM; n = 3 independent homogenation/qRT-PCR experiments; *P < 0.05, **P < 0.01 compared to rd10 vehicle control. b Retinal homogenates were used for determination of MCP-1 protein levels by ELISA. Quantification: normalization to WT mice injected with vehicle; mean ± SEM; n = 3 experiments; *P < 0.05. c Retinal microglia were purified from dissociated rd10 retinas via flow sorting, as described in “Methods”. Sorting was performed five times using retinas collected from five groups of rd10 mice (four mice each). Sorted microglial cells (CD11b + CD45low) were then subjected to qRT-PCR. Quantification: normalization to GAPDH mRNA; mean ± SEM; n = 5; *P < 0.05, **P < 0.01, compared to rd10 vehicle control. d Microglial cells were isolated and purified from B6 mouse brains. Cells were pre-incubated with vehicle (DMSO) or JQ1 (0.5 μM) for 12 h, and then induced for inflammatory cytokine expression with LPS (1 μg/ml) for another 2 h prior to qRT-PCR assay. Quantification: normalization to GAPDH mRNA; mean ± SEM; n = 3 independent experiments; *P < 0.05, **P < 0.01 compared to vehicle + LPS