Fig 2

(A) TALEN generated 17-bp deletion leads to a frameshift mutation resulting in a truncated 27 amino acid (aa) Rad21l1 protein with the conserved Rec8/Rad21-like family domains (1–100 aa and 495–543 aa) disrupted or deleted. Rec8/Rad21-like domains (purple boxes); altered amino acid sequence (red box). The ATG translational start site is located at the 4th-6th nucleotide from the end. (B) Spermatocyte nuclear spreads stained for telomeres (cyan), Sycp3 (green), and Rad21l1 (magenta). Rad21l1 forms lines of foci along the Sycp3 axis in rad21l1^+/- spermatocytes. In the rad21l1 mutant, no lines of Rad21l1 foci are seen. The rad21l1 mutant spermatocytes can form axes and pair homologs albeit with some asynapsed regions. Scale bar = 5 μm. (C) Sexed offspring of a rad21l1^+/- incross show a depletion of females in rad21l1^-/- fish. Data pooled from multiple crosses. (D) Sections of gonads prepared from 35–36 dpf rad21l1^+/+and rad21l1^+/- (labelled rad21l1⁺) and rad21l1^-/- fish and stained for DNA (gray) and Ddx4 (also known as Vasa; green). At 35–36 dpf, oocytes are present in 10/12 rad21l1^+/- and 6/11 rad21l1^-/- samples. Scale bar = 50 μm. Arrows represent different cell types: (pink- diplotene oocytes, yellow- spermatocytes / sperm; red- premeiotic germ cells). (E) Whole mounts of gonads from 40 and 45 dpf are stained for DNA (gray) and Ddx4 (green). At 40 dpf, oocytes are present in 11/21 rad21l1^+/+ and 1/19 rad21l1^-/- samples. At 45 dpf, oocytes are present in 7/9 rad21l1^+/+ and 1/10 rad21l1^-/- samples. Scale bar = 50 μm. Fisher’s exact test used for all statistical analysis. ns = p>0.05, ** = p<0.01, **** = p<0.0001.