Fig. 4: Hsf5 KO spermatocytes failed to progress through pachytene. | Nature Communications

Fig. 4: Hsf5 KO spermatocytes failed to progress through pachytene.

From: Atypical heat shock transcription factor HSF5 is critical for male meiotic prophase under non-stress conditions

Fig. 4

a Seminiferous tubule sections (4 weeks) were stained as indicated. Scale bar: 25 μm. Yellow arrowhead indicates H1t-positive spermatocyte with abnormal SYCP3 staining. pL: preleptotene, Pa: pachytene, rS: round spermatid. Shown on the right is the quantification of the seminiferous tubules that have H1t + /SYCP3+ cells per the seminiferous tubules that have SYCP3+ spermatocyte cells in Hsf5 +/- and Hsf5 KO testes (Mean with SD). n: the number of animals examined. Statistical significance is shown (p = 0.0051, unpaired two-tailed t-test). b–f Chromosome spreads of Hsf5 +/- and Hsf5 KO spermatocytes (P21) were immunostained as indicated. b Lep: leptotene, Zyg: zygotene, Pac: pachytene, Dip: diplotene. Pac* indicates pachytene spermatocyte with high level of γH2AX signals remained on autosomes. Scale bar: 10 μm. Shown on the right is quantification of stages per total SYCP3+ spermatocytes. n: the number of cells examined. c Hsf5 +/- (n = 60) Hsf5 KO (n = 55). Scale bar: 10 μm. Enlarged images of the XY body are shown on the bottom. Scale bar: 5μm. d  ~ 17.8% of Hsf5 KO pachytene spermatocytes (n = 62) exhibited BRCA1 along autosomes with γH2AX signals, whereas none of Hsf5 +/- pachytene spermatocytes (n = 51) did except for XY chromosome. Scale bar: 10 μm. e The number of DMC1 foci is shown in the scatter plot with median (right). Statistical significance is shown (p < 0.0001, two-sided Mann-Whitney U-test). Lep leptotene, Zyg Zygotene, Pac Pachytene. Scale bar: 10 μm. f The number of MLH1 foci is shown in the scatter plot with median (right). Statistical significance is shown (Mann-Whitney U-test). n: number of spermatocytes examined. Statistical significance is shown (p < 0.0001, two-sided Mann–Whitney U-test). Scale bar: 5 μm. g Seminiferous tubule sections (4 weeks) were subjected to TUNEL assay. Whole testis sections (left, Scale bar: 500 μm) and closeup view of seminiferous tubule sections (middle, Scale bar: 25 μm) are shown. Shown on the right is the quantification of the seminiferous tubules that have TUNEL+ cells per total tubules in Hsf5 +/- (n = 3) and Hsf5 KO (n = 3) testes (Mean with SD). Statistical significance is shown by p-value (p = 0.0244, unpaired two-tailed t-test).

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