Table 1

Primer sequences of target and house-keeping genes"

Fig. 1

Immunohistochemical staining of EPO in kidneys of adult cattle and yaks Positive expression is showed in brown colour. B, C, E, F are localised expression of EPO in the kidney of yak and yellow-cattle, respectively, and its positive expression in renal tubular epithelial cells and peritubular mesenchymal fibroblast-like cells. B, E. Original magnification: 200×, Scale=100 μm; C, F. Original magnification 400×, Scale=50 μm; A, D are negative control, original magnification: 200×, Scale=100 μm. TE. Renal tubular epithelial cells; PI. Renal tubular interstitium"

Fig. 2

Expression of EPO protein and mRNA in kidney of adult cattle and yak A. Expression of EPO protein in kidney of adult cattle and yak; B. Gray value analysis of EPO protein expression in the kidney of adult cattle and yak; C. Relative expression of EPO mRNA in the kidney of adult cattle and yak. * P<0.05 indicates that there is a significant difference between the two groups; *** P<0.001 indicates that the difference between the two groups is extremely significant, the same as below"

Fig. 3

Cell primary culture and determination of growth curve A. Cell morphology at P0; B. Cell morphology at P3; C. Cell morphology at P7; D. Determination of cell growth curve at P3: The abscissa is the measurement time, and the ordinate is the OD value"

Fig. 4

Immunofluorescence identification of yak RIFs(100×) A-C. Cell vimentin immunofluorescence staining showed that the protein was mainly expressed in the cytoplasm; D-F. Cell α-smooth muscle actin immunofluorescence staining, the protein is mainly expressed in the cytoplasm; G-I. Cytokeratin 18 immunofluorescence staining showed that the expression of cytokeratin 18 in renal interstitial fibroblast-like cells was negative; the nucleus was stained with DAPI"

Fig. 5

Regulation of apoptosis-related genes at the mRNA level by IOX3-induced EPO A. Therelative expression of EPO mRNA; B. Therelative expression of Bcl-2 mRNA; C. Therelative expression of Bax mRNA; D. Therelative expression of PCNA mRNA. ** P<0.01 indicates that the difference between the two groups is extremely significant, the same as below"

Fig. 6

Regulation of apoptosis-related genes at the protein level by IOX3-induced EPO A. Western blot gel electrophoresis results; B. EPO protein relative expression; C. Bcl-2 protein relative expression; D. Bax protein relative expression; E. PCNA protein relative expression"

Fig. 7

The mRNA expression levels of apoptosis-related genes after inhibiting the expression of EPO A. The relative expression of EPO mRNA; B. The relative expression of Bcl2 mRNA; C. The relative expression of Bax mRNA; D. The relative expression of PCNA mRNA"

Fig. 8

Protein expression levels of apoptosis-related genes after inhibiting EPO expression A. Western blot gel electrophoresis results; B. EPO protein relative expression; C. Bcl-2 protein relative expression; D. Bax protein relative expression; E. PCNA protein relative expression"

Fig. 9

Proliferation and apoptosis assay of RIFs A1-A3. EDU-stained Control group; B1-B3. EDU-stained LW6 group; C1-C3. EDU-stained IOX3 group; D. Flow cytometric Control group; E. Flow cytometric LW6 group; F. Flow cytometric IOX3 group"