Fig. 3: Ex vivo evaluation of seminal plasma toxicity.

a Representative histology (H&E staining) of the explant before and after 2 and 4 h of culture with or without 50% LSP, showing intact epithelium. Magnification ×20. Scale bar = 50 µm. b, c Immunofluorescence staining for the human epithelial antigen (HEA-FITC, green), E-cadherin (red), and the junctional adhesion protein (JAM, green) of explants before and after treatment for 2 h with culture medium (negative control), SIVmac251 with or without 50% LSP, and 50% LSP. Scale bar = 50 µm in (b) and 100 µm in (c). d Integrity of the epithelial barrier measured by the addition of Dextran-FITC (FD4, 4 kDa, 250 µg/ml) to the apical side of the explant with or without 25% or 50% LSP for 2 and 4 h. Results are shown as the percentage of positive control (dye added to the basal medium at the beginning of the experiment). EDTA (100 mM) was included as a control of barrier disruption. The statistical significance between conditions was tested using Wilcoxon signed-rank tests, ***p < 0.0005. e Viability of the explants exposed, or not, for 2 and 4 h to 25% or 50% LSP measured by the MTT assay. A solution of 10% bleach was included as a control for death. Results are shown as the mean ± SD of triplicates from a representative experiment of three.