Figure 3

Tia1 knockout exacerbates the P42 C ^+/+ testis phenotype. (A) Male reproductive organs, including testis, epididymis, vas deferens and seminal vesicle, from P42 mice. Scale bar is 10 mm. (B) Gross testes mass for P42 mice (n = 10, 8, 8 and 7 WT, Tia1 ^−/−, C ^+/+ and C ^+/+/Tia1 ^−/− mice, respectively). (C) Relative testes mass for P42 mice as determined by dividing the testis mass by total BW. Numbers of mice the same as in (B). (D) Serum testosterone level for P42 mice (n = 4 mice per genotype). (E) Representative H&E-stained cross sections from the testis and the cauda epididymis. Genotypes are indicated above each micrograph. Examples of vacuolization (black arrowheads), multinucleated bodies (red arrowheads) and sloughed tissue (blue arrowheads) are marked. Scale bar is 100 µm. (F) Seminiferous tubule health was assessed with a 10-point scale (see Materials and Methods). The graph indicates the average score for each genotype. Numbers of mice are the same as in (B). (G) Cumulative frequency plot of seminiferous tubule scores for all mice. (H) Total epididymal sperm count from P60 mice (n = 8, 12, 6 and 8 WT, Tia1 ^−/−, C ^+/+ and C ^+/+/Tia1 ^−/− mice, respectively). (I) Percentage of seminiferous tubules with at least one TUNEL-positive object as assessed by TUNEL staining. A minimum of 50 seminiferous tubules per mouse were examined (n = 4 mice per genotype). (J) Average number of TUNEL-positive objects per seminiferous tubule (n = 4 mice per genotype). (K) Representative western blots for SMN, Gemin2, Tia1, Tiar and loading control β-actin in P42 testis. Full-length blots are presented in Supplementary Fig. 1. Genotypes and lane numbers are indicated above and below the blots, respectively. The graphs to the right indicate the results of densitometry (n = 4 mice for each genotype). Error bars on all graphs indicate S.E.M. For (B,C,D,F,H,I and J) and (K), statistical comparison between genotypes was performed using a one-way ANOVA with Tukey’s multiple comparison test and the results are denoted by lowercase letters. Bars with different letters indicate a statistically significant different between the means, whereas bars with the same letters are not significantly different (p < 0.05). For (B,C,D,F,H,I and J), data from WT and C ^+/+ mice was previously published³³.