Roles Played by IL-8 in Altering Dynamics of Trabecular Meshwork Cells after Human Cytomegalovirus Infection

RNA-seq analysis

Human trabecular meshwork cells (HTMC) were infected by human cytomegalovirus (CMV). For blocking experiments, HTMC were infected by TB40/E strain of CMV and exposed to CXCR2 or CCR2 antagonists. For blocking CXCR2, an IL-8 receptor, 500 nM of SB225002 (CXCR2 antagonist, Abcam, Cambridge, UK) was used (CMV_SB). For blocking CCR2, 5 µM RS504393 (CCR2 antagonist, Tocris, Bristol, UK) was used (CMV_RS).

Total RNA was isolated from CMV-infected human trabecular meshwork cells. An RNA library was prepared using the Ion AmpliSeq Transcriptome Human Gene Expression Kit (Thermo Fisher Scientific, Waltham, MA), and sequenced using the Ion Proton System (Thermo Fisher Scientific).

The mapped sequencing read data as BAM files were analyzed using featureCounts. Differential expression was analyzed using edgeR.