Fig. 5: The changes in expression pattern showed the heterogeneity and maturation arrest of iNOA Sertoli cells.

a, b UMAP plot showing the heterogeneity among iNOA Sertoli cells; a clear difference is observed between iNOA and normal adult Sertoli cells. c Heatmap of DEGs between healthy adult and iNOA Sertoli cells. d The percentage of overlap genes between iNOA (cluster 1 and cluster 2) and normal developmental Sertoli cells in Stage_a or Stage_b, and between normal adult (cluster 3) and other normal Sertoli cells in Stage_c. e Violin plot of the expression levels of candidate stage-specific markers in three Sertoli cell clusters. The upper row shows Stage_a markers; the middle row shows Stage_b markers, and the bottom row shows Stage_c markers. The larger black dots in each violin plot column represent the mean value. f, g GSEA terms enriched in f cluster 1 and g cluster 2 iNOA Sertoli cells, shown as barplot. The GSEA score is presented on the x-axis, and the gradient of red indicates low to high FDR values. h iNOA Sertoli cells proliferated faster than normal adult Sertoli cells in vitro, as shown by CCK-8 assay. Data shown as mean ± SD from three biological independent samples and two times independent experiments, statistical analysis made by two-tailed, Student’s t-test; the confidence interval is 95%. *p < 0.05 (comparing with normal adult). i Violin plot of the expression levels of energy metabolism-related genes in three Sertoli cell clusters. The larger black dots in each violin plot column represent the mean value. j, k Pathways terms of IPA enriched in j cluster 1 and k cluster 2 compared with normal adult Sertoli cells, shown as barplot. The p-value is presented on the x-axis, and the gradient of two colors indicates low to high activation scores in the two clusters.