Figure 5.

Multilineage interactions in seminiferous tubules. A) Network plots showing ligand–receptor interaction events between different cell types in the four idiopathic nonobstructive azoospermia (iNOA) classes. Cell–cell interactions are indicated by the connecting lines. Line thickness is proportionate with the number of interaction events. Node size represents interaction strength. B) Top 10 ligand–receptor pairs of cell types with the most variation in the number of interactions. C) Heatmaps showing the changes in ligand–receptor interaction events in each class of iNOA compared to the normal controls (NC) group. Red and blue indicate an increase or decrease, respectively, in the number of interactions compared to the NC group. D) Network plots showing the changes in ligand–receptor interaction events between Sertoli cells (ST) and germ cells in each class of iNOA compared to the NC group. Cell–cell interactions are indicated by the connecting lines. Line thickness is proportionate with the number of interaction events. Arrows indicate the direction of the interaction. Red and blue indicate an increase or decrease, respectively, in the number of interactions compared to the NC group. (E) Interaction of ST with spermatogonial stem cells (SSC). Ligands (front) were expressed on ST and receptors (back) were expressed on SSC. Dot size represents significance, defined as −log₁₀ (p‐value). Color bars from blue to red represent the normalized expression values of both ligands and receptors. F) Immunofluorescence of LGR4, RSPO3, and the Sertoli cell marker WT1 in NC and iNOA. The scale bar represents 20 µm. Immunoblotting of LGR4 in testicular tissue of NC and iNOA. Quantification of protein levels is shown in the bottom section. Three samples from each group were analyzed. The two‐tailed Student's t test was used for significance test.