Igg1 control

piPSC-RPE cells were seeded in triplicate on laminin-coated wells in a 96-well plate and allowed to grow to confluence for 3 weeks. Cells were pre-incubated with or without 50 μg/mL anti-integrin αVβ5 (Abcam, Cambridge, United Kingdom) or 50 μg/mL IgG1 control (Abcam, Cambridge, United Kingdom) for 45 minutes, then the medium was replaced and cells were incubated with FITC-labeled ROS (at concentrations as stated above) with or without 50 μg/mL anti-integrin αVβ5, or 50 μg/mL IgG1 control for 2.5 hours at 37°C. Wells were then vigorously washed 3-times with PBS to remove unbound ROS. Fluorescent images were obtained randomly from several fields in the DAPI, FITC wavelength for statistical analysis. Immunocytochemistry of RPE cell marker ZO-1 was performed to visualize cell borders after taking the images.