Fig 2: Expression of MIF in H1 and H9 human embryonic stem cell (ESC) lines.A. qRT-PCR analysis of MIF in H1 and H9 human ESCs, P> 0.05, n = 3; B. The expression of MIF in H1 and H9 human ESCs as detected by ICF; the nucleus is blue, and the depth of green fluorescence indicates the expression level of MIF in ESCs, the deeper the fluorescence color, the denser the distribution and the stronger the expression of the target protein, with a magnification of 1200 and a scale of 10 µm; C. The expression of MIF in H1 and H9 human ESCs as shown by Western blot; D. Semi-quantitative analysis of protein expression, compared with Tubulin, **P <0.01, n = 3. E. MIF content in cell medium of H1 and H9 human ESCs at different times. Compared with H1, P> 0.05. Compared with 24 h, ***P <0.001, ****P <0.0001.

Fig 3: Expression of MIF-related receptors in H1 and H9 cells.A. The expression of MIF-related receptors in human H1 and H9 embryonic stem cells was detected by ICF; DAPI stained nuclei showed blue fluorescence, where the depth of red fluorescence indicates the MIF-related receptors CD74, CD4, CD44, CXCR2, CXCR4 and CXCR7, the deeper the red, the denser the distribution, indicating the higher the related receptor expression level, with a magnification of 1200 and a scale of 10m; B. Western blot was used to detect the expression of MIF-related receptors in human H1 and H9 embryonic stem cells; C. Semi-quantitative analysis of protein expression, compared with Tubulin, ****P <0.0001, n = 3.

Fig 4: Interaction of MIF with its receptors CXCR2 and CXCR7.A. MIF antibody was used as bait protein, and the IB result showed that CXCR7 was successfully precipitated; B. MIF antibody was used as bait protein, and the IB result showed that CXCR2 could not be successfully precipitated; C. CXCR7 antibody was used as bait protein to reverse MIF precipitation, and the IB results showed that MIF was successfully precipitated. IB was the target protein as detected by Western blot, IP was immunoprecipitation (using relevant immunoprecipitation antibody), IgG was the negative control group, and Input was the positive control group. D. Nucleus (blue fluorescence), MIF (green fluorescence), CXCR7 (red fluorescence), magnification of 1200, scale of 10m; E. Nucleus (blue fluorescence), MIF (green fluorescence) and CXCR7 (red fluorescence) image composite, fluorescence quantitative analysis (white tip).