CUT & Tag analysis of primary human retinal microvascular endothelial cells
CUT & Tag analysis was conducted using primary human retinal microvascular endothelial cells treated with 0 or 100ng/ml VEGF. Overall design: CUT& Tag was performed using the Hyperactive Universal CUT& Tag Assay Kit for Illumina (Vazyme). Briefly, Human retinal microvascular endothelial cells (HRMECs) were treated with 0 or 100 ng/ml VEGF for 24 h (Control and VEGF group respectively). Then the HRMECs were bound to concanavalin A-coated beads. After permeabilization and incubation with primary antibody (anti-H3K9la, PTM BIO, PTM-1419RM), the DNA was precisely tagmented with pA-Tn5 transposase and enriched by PCR to create sequencing-ready libraries. The library preparations were sequenced using the Illumina Novaseq platform and 150 bp paired-end reads were generated.