Figure 5.

LOVA could promote the proliferation of undifferentiated spermatogonia and protect undifferentiated spermatogonias from apoptosis damage in vivo

(A) Schematic outline of LOVA injection after 10 days of busulfan treatment in order to simulate reparation after chemotherapy.

(B) Histogram calculation of the proportion of undifferentiated spermatogonia (ZBTB16⁺ cells) compared with Sertoli cells (SOX9⁺ cells) in (A) from six independent experiments. Error bars indicate mean ± SEM, ^∗∗p ≤ 0.01, ns, non-significant.

(C) Histogram calculation of the proportion of cleaved caspase-3⁺ ZBTB16⁺-double labeled apoptotic undifferentiated spermatogonia compared with ZBTB16⁺-labeled undifferentiated spermatogonia in (A) from six independent experiments. Error bars indicate mean ± SEM, ^∗∗∗p ≤ 0.001, ^∗∗∗∗p ≤ 0.0001.

(D) Schematic outline of mouse treated by busulfan simultaneously with LOVA injection in order to estimate the prevention of chemotherapy injury.

(E) Immunofluorescent staining of undifferentiated spermatogonia marker (ZBTB16) and Sertoli cell marker (SOX9) or apoptotic cell markers (cleaved caspase-3) in mouse testis under the treatment of busulfan with or without LOVA injection. Scale bars, 50 μm.

(F) Histogram calculation of the proportion of undifferentiated spermatogonia (ZBTB16⁺ cells) compared with Sertoli cells (SOX9⁺ cells) in (E) from six independent experiments. Error bars indicate mean ± SEM, ^∗p ≤ 0.05, ^∗∗p ≤ 0.01.

(G) Histogram calculation of the proportion of cleaved caspase-3⁺ZBTB16⁺-labeled apoptotic undifferentiated spermatogonia compared with total ZBTB16⁺-labeled undifferentiated spermatogonia in (E) from six independent experiments. Error bars indicate mean ± SEM, ^∗∗∗p ≤ 0.001, ^∗∗∗∗p ≤ 0.0001.

See also Figure S4.