For RNA isolation, CNV and surrounding tissues of eyes treated with Fc, VEGF-Trap, and ABTAA were used. Total RNA was isolated using TRIzol reagent (Invitrogen). RNA quality was assessed by an Agilent 2100 bioanalyzer using the RNA 6000 Nano Chip (Agilent Technologies), and RNA quantification was performed using the ND-2000 Spectrophotometer (Thermo Fisher Scientific). For library preparation, the SENSE 3′ mRNA-seq Library Prep Kit (Lexogen Inc.) were used according to the manufacturer’s instructions. Briefly, each 500 ng of total RNA was prepared, oligothymidilate primers containing an Illumina-compatible sequence at the 5′ end were hybridized to the RNA, and reverse transcription was performed. After degradation of the RNA template, second-strand synthesis was initiated by a random primer containing an Illumina-compatible linker sequence at its 5′ end. The double-stranded library was purified by using magnetic beads to remove all reaction components. The library was amplified to add the complete adapter sequences required for cluster generation. The finished library was purified of PCR components. High-throughput sequencing was performed as single-end 75 sequencing using NextSeq 500 (Illumina Inc.). For RNA-seq, the SENSE 3′ mRNA-seq reads were aligned using Bowtie2 version 2.1.0 (52). Bowtie2 indices were either generated from genome assembly sequence or representative transcript sequences for alignment to the genome and transcriptome. The alignment file was used to assemble transcripts, estimate their abundances, and detect differential expression of genes. DEG was determined on the basis of counts from unique and multiple alignments using EdgeR within R version 3.2.2 (R development Core Team) using BIOCONDUCTOR version 3.0 (53). The RT (read count) data were processed on the basis of Quantile normalization method using the Genowiz version (Ocimum Biosolutions).

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