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. 2023 Feb 10;24(4):3587. doi: 10.3390/ijms24043587

Figure 1.

Figure 1

Arg-ii deficiency in human renal epithelial cells prevents hypoxia-induced paracrine effects of the cells on podocyte cytoskeleton derangement. (A) Immunoblotting revealing arg-ii deficiency in arg-ii−/− HK2 cells. (B) Schematic illustration of the experimental setup to study crosstalk between proximal tubular epithelial cells (PTECs) and podocytes. Human PTECs (HK2-CRISPR-wt and -CRISPR-arg-ii−/−) were exposed to either normoxia (21% O2) or hypoxia (1% O2) conditions for 48 h. Conditioned medium (CM) was then collected from HK2 cells and transferred to the human differentiated podocyte (AB8/13) for 24 h of incubation. Cell lysates of AB8/13 were then prepared and subjected to downstream analysis. (C) Representative images showing phalloidin staining of cytoskeletal actin fibers (green) in human podocytes treated with different HK2-CM as indicated. The nucleus was stained with DAPI (blue). (D) The quantification of the podocytes with disrupted actin cytoskeleton (in percentage). (E) Immunoblotting analysis of the protein levels of β-catenin; GAPDH serves as the loading control. (F) Quantification of the β-catenin signals. * p < 0.05, ** p < 0.01, *** p < 0.001 between the indicated groups. n = 3; CM: conditioned media, N: normoxia, H: hypoxia.