ANTXR1 Regulates Erythroid Cell Proliferation and Differentiation through wnt/β-Catenin Signaling Pathway In Vitro and in Hematopoietic Stem Cell

Figure 6

The Wnt/β-catenin signaling pathway cooperates with ANTXR1 in regulating erythroid cell differentiation. (a) The expression of ANTXR1 and LRP6 in K562 cells was detected by immunofluorescence with a confocal laser scanning microscope. ANTXR1 was labeled with 428 nm FITC, which represents green; LRP6 was labeled with Texas red; and nuclei were stained by DAPI to compare to the cell membrane. The merged images indicate the localization of ANTXR1 and LRP6 in the cell membrane, which the overlay of green and red give yellow. (b) The protein levels of β-catenin (92 kDa), P-β-catenin (85 kDa), and GS3β (46 kDa) were measured by western blotting after K562 cells were overexpressed or knocked down by ANTXR1. (c) Respectively, with K562 cells overexpressing ANTXR1 treated with 2.5, 5, and 10 μmol/L of XAV939 for 24 h. Cell proliferation was determined by CCK-8. (d) K562 cells knockdown ANTXR1 treated with 20 mM LiCl for 24 h, and cell proliferation was determined by CCK-8. (e) Cord blood CD34⁺ cells knockdown ANTXR1 treated with 5, 20, and 40 mM LiCl for 24 h, respectively. Flow cytometry detection of CD71 and CD235a expression. Error bars represent standard deviation of three independent experiments. , .

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