Fig. 5: Assessment of the effect of in vivo per- and polyfluoroalkyl substances (PFAS) exposure on sperm cell integrity.

Mature spermatozoa were isolated from the cauda epididymides of mice in each exposure group before being immediately assessed for A viability using a Sytox Green viability stain, and B total and C progressive motility using computer assisted sperm analysis (CASA). Alternatively, the cells were subjected to a D MitoSOX Red (MSR) assay to determine the level of mitochondrial reactive oxygen species (ROS) generation. Genome integrity was assessed using a combination of the E sperm chromatin structure assay (SCSA) to determine DNA fragmentation and F an alkaline Comet assay to detect DNA strand breakage. Data are presented as mean ± SEM, calculated on the basis of n = 12 biological replicates for motility, n = 18 for viability and MSR, and n = 4 biological replicates for SCSA, and Comet. All data were subjected to one-way ANOVA with Tukey’s multiple comparisons test.