RNA-seq of rd1 retinal microglia after treatment with AAV8-GFP vs. AAV8-GFP + AAV8-TGFB1

Transcriptional profiling of microglia (seven biological replicates per experimental condition) or non-microglia (four biological replicates total). One thousand microglia (CD11b+ Ly6G/Ly6C-) or non-microglia cells (CD11b-) from P30 rd1 (FVB) retinas infected with either AAV8-GFP or AAV8-GFP + AAV8-TGFB1 were sorted into 10 µL of Buffer TCL (Qiagen) containing 1% beta-mercaptoethanol and immediately frozen on dry ice. Samples were subsequently sent to the Broad Institute Genomics Platform for cDNA library synthesis and sequencing using a modified Smart-Seq2 protocol with an expected coverage of ~6 million reads per sample. Prior to analysis, reads were subjected to quality control measures and mapped to the GRCm38.p6 reference genome. Reads assigned to each gene were then quantified using featureCounts and normalized and analyzed for differential expression using DESeq2. RNA-seq of microglia and non-microglia from P30 rd1 (FVB) retinas treated with AAV8-GFP vs. AAV8-GFP + AAV8-TGFB1