Fig. 2

Deletion of the Tug1 locus leads to sperm defects and male infertility. a Deletion strategy of the Tug1 locus (shown inverted). The Tug1 gene body was replaced by a lacZ reporter cassette, leaving the promoter and first exon intact. The dashed lines indicate the deleted region in the Tug1 knockout. RNA sequencing (RNA-seq) tracks for wild-type (WT) and Tug1^−/− testis are depicted. b Scatter dot plot (showing the mean and standard deviation) of the number of pups at birth per copulatory plug for matings between wild-type, Tug1^+/−, or Tug1^−/− males and wild-type C57BL/6J females (left panel) and wild-type C57BL/6J males and wild-type, Tug1^+/−, or Tug1^−/− females (right panel). Each dot represents a litter from a different mouse. Significant (*) p value (Wilcoxon rank-sum test with Bonferroni correction) and the number of mice for each genotype tested are indicated. c Box plot of total sperm count for wild type and Tug1^−/− males. Significant (*) p value (Wilcoxon rank-sum test) is indicated. d Box plots of the percentage of normal sperm and sperm with the five most common morphological abnormalities for wild-type (n = 9) and Tug1^−/− (n = 8) males. Representative images of normal and morphologically aberrant sperm are located below each corresponding plot. Red arrows indicate the location of the defect. Scale bars are 20 μm. Significant (*) p values (Wilcoxon rank-sum test) are indicated. e Representative spermatocyte diagrams and micrographs of Tug1^+/− seminiferous tubule sections stained with periodic acid-Schiff’s reagent and X-gal showing expression of the lacZ reporter under the control of the endogenous Tug1 promoter at the indicated stages of spermatogenesis. Scale bars are 20 μm. f Representative spermatid diagrams and micrographs of wild-type and Tug1^−/− epididymis tubule sections stained with hematoxylin and eosin. Scale bars are 20 μm