Fig. 3: PRRC2A deficiency leads to chromosome misalignment and spindle disorganization.

a Immunostaining of pH3 and γH2AX in P60 control and Prrc2a-cko testis sections. Seminiferous tubule stages are indicated. Arrowheads indicate metaphase spermatocytes. Scale bar, 50 μm. b The number of pH3⁺ spermatocytes per seminiferous tubule in control and Prrc2a-cko testes of indicated ages (n = 4 mice for P18, P20, P60; n = 3 mice for P22, P24, P28, P35). Two-sided student’s t-test. Error bars, mean ± SEM. p = 0.7785 (for P18), 0.0149 (for P20), 0.0334 (for P22), 0.0231 (for P24), 0.0250 (for P28), 0.6855 (for P35), 0.4705 (for P60). Source data are provided as a Source Data file. c The ratio of seminiferous tubules with pH3⁺ spermatocytes to total tubules in control and Prrc2a-cko testes of indicated ages (n = 4 mice for P18, P20, P60; n = 3 mice for P22, P24, P28, P35). Two-sided student’s t-test. Error bars, mean ± SEM. p = 0.7580 (for P18), 0.0014 (for P20), 0.1047 (for P22), 0.1941 (for P24), 0.0092 (for P28), 0.0456 (for P35), 0.0054 (for P60). Source data are provided as a Source Data file. d The number of pH3⁺ spermatocytes per seminiferous tubules of the indicated stage in P60 control and Prrc2a-cko testes. Two-sided student’s t-test. Error bars, n = 4 mice, mean ± SEM. p = 0.0014 (for I–III), 0.0591 (for IV–VII), 0.0242 (for VIII–X), 0.0176 (for XI–XII). Source data are provided as a Source Data file. e H&E staining in P60 control and Prrc2a-cko testes sections. Arrowheads indicate misaligned chromosomes in metaphase I spermatocytes. Scale bar, 20 μm. f Representative metaphase I spermatocytes in P60 control and Prrc2a-cko testis sections immunostained with α-tubulin and γ-tubulin. White arrowheads indicate misaligned chromosomes, yellow arrowheads indicate scattered γ-tubulin foci outside the spindle pole, yellow arrows indicate disintegrated MTOC at the spindle pole, white arrows indicate spindle poles. Scale bar, 5 μm. g Percentage of metaphase I spermatocytes with chromosomes misalignment (n = 4 biological independent mice), scattered γ-tubulin foci, disintegrated MTOC, and disorganized spindle (n = 3 biological independent mice) in P60 control and Prrc2a-cko testes. Two-sided student’s t-test. Error bars, mean ± SEM. p = 0.0007, 0.0018, 0.0423, 0.0057. Source data are provided as a Source Data file. h Fluorescence intensity of γ-tubulin at the spindle pole relative to that in control (n = 97, 100 cells). Two-sided student’s t-test. Error bars, mean ± SEM. ***p < 0.0001. Source data are provided as a Source Data file. i, j The average length and width of the alignment and spindles in control and Prrc2a-cko metaphase I spermatocytes immunostained with α-tubulin and γ-tubulin (n = 61 cells for control, n = 64 cells for Prrc2a-cko). Two-sided student’s t-test. Error bars, mean ± SEM. The value of each data is 10.21 ± 0.16, 10.07 ± 0.13, 6.07 ± 0.15, 7.55 ± 0.15, 9.05 ± 0.32, 10.25 ± 0.22, 7.25 ± 0.16, 7.38 ± 0.15 from left to right. p = 0.5051 (ns), <0.0001 (***), 0.0018 (**), 0.5759 (ns). Scale bar, 5 μm. Source data are provided as a Source Data file.