Additional file 6 of Single-cell transcriptome analysis and in vitro differentiation of testicular cells reveal novel insights into male sterility of the interspecific hybrid cattle-yak
Creators: name: Mipam, TserangDonko
name: Chen, Xuemei
name: Zhao, Wangsheng
name: Zhang, Peng
name: Chai, Zhixin
name: Yue, Binglin
name: Luo, Hui
name: Wang, Jikun
name: Wang, Haibo
name: Wu, Zhijuan
name: Wang, Jiabo
name: Wang, Mingxiu
name: Wang, Hui
name: Zhang, Ming
name: Wang, Hongying
name: Jing, Kemin
name: Zhong, Jincheng
name: Cai, Xin
Publisher: name: figshare
Year: 2023
Subjects: subject: Genetics
subject: FOS: Biological sciences

description: Additional file 6: Fig. S1. Identification of pubertal testicular cells from cattleyak and yak. (A and B) The re-suspension cells of cattleyak and yak after 2 days of culture, respectively. (C and D) The size and distribution of testicular cells from cattleyak and yak, respectively. (E and F) Immunofluorescence of DDX4 (green) in pubertal testicular cells of cattleyak and yak, respectively. The scale bars represent 50 μm. Fig. S2. Validation of cell type specific gene expressions between testicular cells of cattleyak and yak. (A) Log10 fold change values for RT-qPCR detection of 46 germ cell specific signature gene expression in pubertal testes between cattleyak and yak. (B) Log10 fold change values for RT-qPCR detection of 10 niche cell type specific signature gene expression in pubertal testes between cattleyak and yak. YK and CY denote yak and cattleyak, respectively. * indicates the differentially expressed signature genes in undifferentiated spermatogonial cells. Fig. S3. Dot plot showing relative expression patterns of the potential marker genes for each germ cell type (A) Relative expression of the potential marker genes for spermatogenic cells in cattleyak. (B) Relative expression of the potential marker genes for spermatogenic cells in yak. Fig. S4. Proliferation analysis of the spermatogenic cells cultured in vitro by EDU staining. (A, B) EDU and DAPI staining of cattle-yak and yak spermatogenic cells, respectively. Fig. S5. Spermatogenic cells co-cultured with different testis somatic cells and cell viability analysis. (A) Yak spermatogenic cells co-cultured with yak testicular somatic cells as feeder and sub-cultured three times in vitro. (B) Cattleyak spermatogenic cells co-cultured with cattleyak testicular somatic cells as feeder and sub-cultured three times in vitro. (C) Cattleyak spermatogenic cells co-cultured with yak testicular somatic cells as feeder and sub-cultured three times in vitro. (D) CCK8 assay detected the viability of yak and cattleyak spermatogenic cells co-cultured with their testicular somatic cells for 4 consecutive days. The scale bars represent 50 μm. * denotes P&lt;0.05, and *** denotes P&lt;0.001. Fig. S6. Inducion of spermatogenic cells of cattleyak and yak with retinoic acid (RA) in vitro. (A) STR8, c-kit, BOLL, DAZL and SCYP3 mRNA expressions in of spermatogenic cells of cattleyak and yak after AR inducion. (B and C) The cell size and distribution of spermatogenic cells after AR induction in yak and cattle yak, respectively. (D) Percentage of spermatocytes of spermatogenic cells after AR inducion in yak and cattle yak, respectively. (E) SYCP3 protein expression in spermatogenic cells after AR induction in yak and cattleyak, respectively.