Fig 3. Mutations of tsn result in abnormal spermatogenesis.

(A) The EMS mutant alleles of tsn. The position of each mutation is indicated by an arrow above a schematic representation of the TSN protein. tsn⁰⁵³⁶ and tsn⁰⁶¹⁴ are point mutations at 282Ile→Phe and 320Glu→Lys of the SN2 domain, respectively. tsn⁰²⁵¹ and tsn¹²³⁶ are point mutations at 368Gly→Glu and 501Gly→Arg of the SN3 domain, respectively. tsn¹¹²⁰ is a deletion mutation which causes frameshift from 437Ile of the SN3 domain. (B) TSN protein expression level in the adult testes of each mutant allele heterozygous with a deficiency (Df) that disrupts the TSN genomic locus was analyzed by Western blots. TSN was undetectable in the lysates from tsn¹¹²⁰/Df testes. (C-H) Immunostaining of TSN (using mouse anti-TSN antibody, green) and VASA (red) in the testes from the five tsn mutant alleles heterozygous with the Df allele. DNA was labeled by DAPI (blue). tsn⁰⁶¹⁴/Df (F), tsn¹¹²⁰/Df (G), and tsn¹²³⁶/Df (H) resulted in the swollen apical tip of the testis. TSN was barely detectable in the tsn¹¹²⁰/Df testis (G). tsn⁰²⁵¹/Df (D) and tsn⁰⁵³⁶/Df (E) did not cause obvious testicular phenotype compared to the WT testis (C).