Figure 1.

In vivo microinjection and electroporation of mouse testes

(A) Macroscopic appearance of a testis transfected with reporter plasmids. Microinjection involved efferent duct injection and filled 70%–80% of the tubules in each recipient testis. (B–E) The localization of GFP protein in testis. Immunofluorescence analysis of GFP localization in testis after electroporation. Promyelocytic leukemia zinc finger (PLZF) and SYCP3 served as markers of SSCs and spermatocyte cells, respectively. PNA is an established marker of the mammalian acrosome, and WT1 served as a marker for Sertoli cells. The white arrowhead indicates GFP signal in elongated spermatids. Scale bar, 2.5 μm. (F and G) Western blot analysis of testis extracts after electroporation. GAPDH and actin served as loading controls. (H) Western blot analysis of testis extracts after different times of electroporation. Actin served as a loading control.