Figure 1
Expression of CXCR3 in mouse testicular cells. (a) Identification of testicular cells. Major testicular cells, including Leydig cells (LC), Sertoli cells (SC) and germ cells (GC), were isolated from 4-week-old C57BL/6J mice. Each cell type was identified by immunostaining for respective markers: LHR for LC, WT1 for SC and MVH for GC. Insets in the upper right corners of the images represent negative controls, in which the preimmune rabbit sera were used as the first antibodies. (b) CXCR3 mRNA. Cells were cultured in the absence (Ctrl) and presence of 107 PFU/ml MuV for 24 h. Total RNA was extracted from the testicular cells, and relative mRNA level of CXCR3 was determined using real-time qRT-PCR by normalizing to β-Actin. The lowest CXCR3 mRNA level in SC was set as ‘1’. The fold increases in LC and GC compared to SC were presented. (c) CXCR3 protein. Testicular cell lysates were subjected to western blot analysis to probe CXCR3 using specific antibodies. β-Actin was probed as loading controls. (d) CXCR3 distribution in the testis. Immunohistochemistry was performed to localize CXCR3 on the paraffin sections of the testis from 5-week-old C57BL/6J mice. The inset in the upper right corner of the image in the right panel represents negative control, in which the preimmune goat serum was used as the first antibody. Black arrows, black arrowheads, white arrows, white arrowheads and asterisk indicate round spermatids, SC, spermatogonia, spermatocytes and interstitial cells, respectively. (e) CXCR3 locations in GC and SC co-cultures. IF co-staining with CXCR3 (green) and MVH (red) for GC and SC co-cultures. Scale bar=20 μm. Images are the representatives of at least three experiments. Real-time qRT-PCR data are the means±S.E.M. of three experiments
