Fig. 9: Damaged epididymis and impaired fertility of 8 weeks DT-injected Foxp3-DTR mice.

H&E staining of the proximal (IS and caput) (A) and distal (corpus and cauda) (B) epididymal regions of DT-injected WT and Foxp3-DTR mice and quantification of the number of damaged sites normalized per tissue area (µm²). Any morphological alteration in epididymal epithelia (arrows) was counted as a damaged site. Each area quantified is represented as a dot. A: n = 9 pictures from 3 prox. epididymides from different mice; B: n = 10 (WT); n = 8 (Foxp3-DTR) images from 4 dist. epididymides from different mice. Computer-assisted sperm analysis (CASA) analysis of proximal sperm cells (C) and distal sperm (D) counts. n = 6 (WT), n = 11 (Foxp3-DTR). E Percentage of total sperm motility analyzed by CASA of distal sperm. n = 6 (WT), n = 11 (Foxp3-DTR). F Fertility assessment by natural mating as the number of litter size (pups/litter). n = 7 (WT), n = 10 (Foxp3-DTR). G Graphical representation of the key findings at 2 and 8 weeks after Treg depletion in the epididymis. The figure was created with BioRender.com. L: Lumen. Bars: 250 μm. Data were analyzed using two-sided Student’s t-test (C–E) or Mann–Whitney test (A, B, F). Data are shown as means ± SEM. n = number of samples from different mice.