Activated Müller glia-conditioned media possesses antibacterial properties. MIO-M1 cells were left either untreated or infected with S. aureus (SA) for four hours, the respective ‘normal’ (untreated), ‘activated’ (SA challenged), and proteinase K (20µg/ml for 30 minutes) treated culture media were collected. In a separate set of wells, bacteria were cultured in DMEM media for four hours without cells. All conditioned media were passed through a 0.22 μm syringe filters (bacteria/cell free) and applied to cover slips containing SA biofilms for four hours (A). Bacterial killing was assessed via Live/Dead® staining and subsequent fluorescent microscopy (B). In comparision to untreated, bacterial growth, and proteinase K treated conditioned media, the activated conditioned medium exhibited antibacterial properties, as evidenced by the increased red staining. The experiment was repeated three times independently and the representative Figure is shown.