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Additional file 5: Table S5. Primer sequences for detection of germ cell marker genes during spermatogenic cell proliferation and differentiation between cattle-yak and yak.

Additional file 4: Table S4. Primer sequences for validation of cell type specific gene expressions in testicular cells between cattle-yak and yak.

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

Additional file 1: Table S1. Statistics of cell numbers, scRNA sequencing and transcriptome alignment.

Peroxisome proliferator-activated receptor-gamma (PPARG) is considered to be a central regulator of lipid metabolism in mammary cells. Adipose differentiation-related protein (ADRP), a member of PAT family (Perilipin, ADRP, TIP47 family), plays a key role in lipid accumulation in mammary gland. It has been found that PPARG significantly promoted lipid storage in goat mammary epithelial cells (GMEC), which was abolished after knockdown of ADRP gene. The results of real-time PCR, Western blot and luciferase reporter assay for goat ADRP promoter showed that ligand-activated PPARG up-regulated the ADRP gene expression and activity of ADRP promoter. Moreover, PPARG directly interacted with a PPRE (PPAR response element) spanning at -1003 to -990 on ADRP promoter. In this study, to our knowledge, we are the first to verify the function of PPARG on lipid storage on cellular level of goat mammary gland and our results revealed a novel pathway that PPARG may regulate lipid accumulation by controlling the expression of ADRP gene.

Peroxisome proliferator-activated receptor-gamma (PPARG) is considered to be a central regulator of lipid metabolism in mammary cells. Adipose differentiation-related protein (ADRP), a member of PAT family (Perilipin, ADRP, TIP47 family), plays a key role in lipid accumulation in mammary gland. It has been found that PPARG significantly promoted lipid storage in goat mammary epithelial cells (GMEC), which was abolished after knockdown of ADRP gene. The results of real-time PCR, Western blot and luciferase reporter assay for goat ADRP promoter showed that ligand-activated PPARG up-regulated the ADRP gene expression and activity of ADRP promoter. Moreover, PPARG directly interacted with a PPRE (PPAR response element) spanning at -1003 to -990 on ADRP promoter. In this study, to our knowledge, we are the first to verify the function of PPARG on lipid storage on cellular level of goat mammary gland and our results revealed a novel pathway that PPARG may regulate lipid accumulation by controlling the expression of ADRP gene.

A strain of Escherichia coli was positive for 5 aminoglycoside modifying enzyme genes (aac(3)-I, aac(6′)-Ib-cr, ant(3′)-1, aadA5, and aph(3′)-I) in PCR assays. And these positive genes confer resistance to aminoglycosides (gentamicin and tobramycin). This is the first report of emergence of five kinds of aminoglycoside-modifying enzymes genes simultaneously in E. coli worldwide.