Fig. 2: Single-cell analysis of human diabetic testis suggests impaired cell junction in BTB.

a Scatter plot showing the DEGs in Sertoli cells between diabetic patients and normal spermatogenesis. b Enriched GO terms in the upregulated and downregulated genes of Sertoli cells from diabetic versus with normal control. The statistical test is a hypergeometric test. c GSEA analysis showing the upregulated and downregulated pathway activity in Sertoli cells. d Boxplots showing the expression levels of Activation NMDARS, Pyruvate metabolism, P38 MAPK pathway, BTB related genes (Normal = 43 cells, DM = 42 cells). Each box represents the median and the 25% and 75% quartiles, and the whiskers indicate 1.5 times of the interquartile range. e Heatmap showing the representative decreased metabolic pathway activity from the normal control and diabetic data in Sertoli cells. f H&E staining of adult human testicular paraffin sections from donors with normal control (left) and diabetic (right), arrow indicated vacuoles and free cells in seminiferous tubules. Scale bar, 20 μm. g–j Immunofluorescence of GJA1, TJP1, NACM1 and CDH2 (red) co-stained with DDX4 (green) in normal and diabetic patients’ testicular paraffin sections. n = 3 testis samples per group. Representative immunofluorescence images and quantitative analysis of GJA1, TJP1, NACM1 and CDH2. Scale bar, 10 μm. Representative immunofluorescence images and quantitative analysis of CDH2. Scale bar, 10 μm. Fluorescence intensity values of more than 50 positive cells in at least 5 fields of view were counted in normal and diabetic patients’ testicular paraffin sections. Two-tailed student’s t test was performed. 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).