Testicular germ cell-specific elimination of Inv::GFP protein in Inv-KD mice.

<p>Testicular sections prepared from adult (a–f and i–p) or postnatal (3 day) (g and h) Inv-KD mice (b, d, f, h, j, l, n and p) or control Inv::GFP-rescue mice (a, c, e, g, i, k, m and o) were stained with anti-GFP antibodies (g, h, m and n, red) or double-stained with anti-GFP antibodies (c, d, i–l, o and p, red) and testicular germ cell-specific anti-calmegin antibodies (e, f, k and l, green) or vascular endothelial cell-specific PECAM-1 antibodies (o and p, green). Hoechst-stained images are shown to identify the cell types of the testis (a, b, g–i and m–p, blue). Inv::GFP protein was mainly observed in the spermatid cells of testicular germ cells (a, c, e, i and k, arrowhead) and somatic cells, such as connective tissue cells and Leydig cells (m, double arrows and arrow) and vascular endothelial cells (o, arrow) in Inv::GFP-rescue mice. Inv::GFP protein was specifically eliminated in the germ cells of Inv-KD mice (b, d, f, j, and l), and was down-regulated in connective tissue and Leydig cells (n, double arrow and arrow) and vascular endothelial cells (p, arrow). In Inv-KD mice, Inv::GFP protein was eliminated in anti-calmegin antibody-positive germ cells in adult testis, and in spermatogonia cells in postnatal mice (day 3) testis (h, arrowhead) but not in Inv::GFP-rescue mice (g, arrowhead). Scale bars: 200 µm (a and b); 50 µm (g−j and m–p).