Fig 1: Relative protein expression of SOD1 (A), GPX1 (B), and GPX4 (C), determined via immunofluorescent staining in bovine spermatozoa treated with BPA, BPS, and BPF. Images were taken at 40× objective using the Olympus FV1200 confocal microscope at laser wavelengths of 405 nm for Hoechst (blue) and 488 nm for Alexa-Fluor 488 (FITC) (green). Localization of respective antioxidants can be determined based on the location of the Alexa-Fluor 488 present. (D) demonstrates the corrected total cell fluorescence (CTCF) of SOD1, GPX1, and GPX4 in bovine spermatozoa determined using ImageJ software. Fluorescent intensity of Alexa-Fluor 488 correlates with protein expression. No significant differences were detected.