Fig. 5. Targeting APLN/APJ repairs BTB damage and improves sperm quality in diabetic mice.

a Schematic illustration of the APLN or ML221 injection experiment in db/db. b Immunofluorescence of biotin (red) between indicated sample groups. Scale bar, 50 μm. c Biotin positive seminiferous tubules percentage in Control, APLN and ML221 injection group. Data are presented as means ± SEM. One-way ANOVA. Statistics were performed in five mouse testes each group (n = 5). d Immunofluorescence of TJP1 and GJA1 (green) co-stained with VIM (red) in APLN injection and ML221 injection testicular paraffin sections. Scale bar, 10 μm. e Quantitative analysis of TJP1 and GJA1. Box-and-whisker plots denote the maximum (top whisker), 75th (top edge of box), 25th (bottom edge of box) and minimum (bottom whisker) percentiles, and the median (line in box). Statistics were performed in five mouse testes each group (n = 5). One-way ANOVA was performed. f Schematic illustration of the ML221 injection experiment in db/db for IVF and ICSI. Mice were treated with ML221 at a dose of 10 mg per kg body weight per day. g Bright field diagram of testicular size in control and ML221 injection group. Scale bar, 2 mm. h H&E staining of testicular sections in control and ML221 injection group. Scale bar, 100 μm. i Sperm counts, sperm motility and testosterone level between control and ML221 injection group. PR: progressive motile, NP: non-progressive motile, IM: immotility. unpaired two-tailed t test was performed. j Brightfield diagram of 4-cell, morula, and blastocyst between control and ML221 injection group. Arrows indicated normal developing embryos. Scale bar, 200 μm. k The trilinear table shows all the embryo injection, two-cell embryos, blastocysts and live C-section-born mice data of IVF between control and ML221 injection group. l The trilinear table shows all the embryo injection, two-cell embryos, blastocysts and live C-section-born mice data of ICSI between control and ML221 injection group.