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. Author manuscript; available in PMC: 2024 Jul 1.
Published in final edited form as: Andrology. 2022 Nov 8;11(5):789–798. doi: 10.1111/andr.13329

Figure 1: Characterization of PDCL2 and immunostaining of PDCL2, CALMEGIN, GOPC, and IZUMO1 in testicular germ cells.

Figure 1:

(A) The expression of mouse Pdcl2 in various organs examined by RT-PCR. Pdcl2 is testis- and epididymis-enriched. Hprt was used as an expression control. He, heart; Li, liver; Sp, spleen; Lu, lung; Ki, kidney; Br, brain; St, stomach; In, intestine; Te, testis; Ov, ovary; Ut, uterus; Ep, epididymis.

(B) The expression of mouse Pdcl2 on indicated postnatal days in the testis was examined by RT-PCR. Pdcl2 begins expression at postnatal day 10. Hprt was used as an expression control.

(C) The expression of human PDCL2 in various organs examined by RT-PCR. PDCL2 is testis-specific. GAPDH was used as an expression control. Li, liver; Ki, kidney; Br, brain; In, intestine; Ut, uterus; Ep, epididymis; H, head; B, body; T, tail.

(D) PDCL2 expression in wild-type testis (Te), purified testicular germ cells (TGC), and germ cell depleted male mice (W/Wv) was examined by immunoblot analysis. PDCL2 is not expressed in W/Wv mutant testis. GAPDH was used as an expression control.

(E) Immunostaining of PDCL2 in wild-type spermatids. Isomorphic to CALMEGIN, PDCL2 is localized in the ER. CALMEGIN is a TGC-specific marker for the ER. GOPC is a marker for the Golgi. Nuclei were stained with Hoechst 33342 (blue). Scale bars: 5 μm.

(F) Round spermatids were observed at step 5–6. Isomorphic to CALMEGIN, PDCL2 is localized in the ER. PDCL2, CALMEGIN, and GOPC signals were detected (red) at step 5–6 round spermatids. CALMEGIN is a TGC-specific marker for the ER. GOPC is a marker for the Golgi. Nuclei were stained with Hoechst 33342 (blue). Scale bars: 5 μm.

(G) Testicular spermatozoa were observed at step 15–16. PDCL2, CALMEGIN, and GOPC signals disappeared at step 15–16 spermatozoa in mouse testis. CALMEGIN is a TGC-specific marker for the ER. GOPC is a marker for the Golgi. Nuclei were stained with Hoechst 33342 (blue). Scale bars: 5 μm.

(H) Protein expression of PDCL2 in wild-type and Pdcl2−/− testicular germ cells and cauda epididymal spermatozoa. BASIGIN was used as a loading control.