Journal: Annals of Translational Medicine

Article Title: The important roles and molecular mechanisms of annexin A 2 autoantibody in children with nephrotic syndrome

doi: 10.21037/atm-21-3988

Figure Lengend Snippet: Autoantibody against glomerular podocyte annexin A2 protein can be detected in the kidney tissues of children with PNS. (A) IgG in the glomerulus (a,e), Annexin A2 expressed on podocytes (b,f), and nephrin expressed on the podocyte foot process slit diaphragm (c,g) could be locally co-localized in the glomeruli of children with PNS (d,h); (e-h) are enlarged images of the framed areas in (a-d), respectively. Scale bar of (a-d): 100 µm, Scale bar of (e-h): 50 µm. (B) Normal control. (a-d) Scale bar: 50 µm. (C) The eluted IgG reacted with both the Annexin A2 protein band isolated from the kidney tissues and recombinant Annexin A2 protein, and the addition of anti-IgG4 secondary antibody confirmed that the eluted anti-Annexin A2 IgG antibodies included the IgG4 subtype. 1, recombinant Annexin A2 protein; 2, Annexin A2 protein band isolated from total protein of human podocytes after SDS-PAGE. Indirect immunofluorescence method was used for staining.

Article Snippet: The kidney slices were washed twice with phosphate-buffered saline (PBS) for 5 minutes at room temperature, blocked with 3% BSA at room temperature, and then incubated overnight at 4 °C with the primary antibodies for nephrin (0.4 μg/mL; #AF3159, RD), ZO-1 (7 μg/mL; #21773-1-AP, Proteintech), WT1 (4.8 μg/mL; #ab89901, Abcam), and desmin (1.2 μg/mL; 16520-1-AP, Proteintech).

Techniques: Control, Isolation, Recombinant, SDS Page, Immunofluorescence, Staining

Journal: Annals of Translational Medicine

Article Title: The important roles and molecular mechanisms of annexin A 2 autoantibody in children with nephrotic syndrome

doi: 10.21037/atm-21-3988

Figure Lengend Snippet: Intravenous injection of anti-Annexin A2 antibody can cause podocyte injury and proteinuria in BALB/c mice. (A) Proteinuria was observed in BALB/c mice intravenously injected with rabbit polyclonal Annexin A2 antibody. An equal amount of rabbit IgG polyclonal antibody-isotype was injected as a control. (B) Sensitization induced by rabbit polyclonal anti-Annexin A2 antibody was observed in BALB/c mice at 24 hours after intravenous injection of the antibody, whereas no IgG antibodies were detected in the glomeruli of mice intravenously injected with an equal amount of rabbit IgG polyclonal antibody-isotype control. The small box in the lower left corner shows the western blotting results for the antibody eluted from the mouse glomeruli after reaction with recombinant Annexin A2 protein. Scale bar: 50 µm. (C) Intravenous injection of anti-Annexin A2 antibody into BALB/c mice led to the downregulation of nephrin, WT-1, and ZO-1, and the upregulation of desmin in the glomerulus. All P<0.05. Scale bar: 50 µm. Indirect immunofluorescence method was used for staining.

Article Snippet: The kidney slices were washed twice with phosphate-buffered saline (PBS) for 5 minutes at room temperature, blocked with 3% BSA at room temperature, and then incubated overnight at 4 °C with the primary antibodies for nephrin (0.4 μg/mL; #AF3159, RD), ZO-1 (7 μg/mL; #21773-1-AP, Proteintech), WT1 (4.8 μg/mL; #ab89901, Abcam), and desmin (1.2 μg/mL; 16520-1-AP, Proteintech).

Techniques: Injection, Control, Western Blot, Recombinant, Immunofluorescence, Staining