Fig. 1

The antigen specificity of anti-PLA2R antibodies. HC group: healthy controls; Ab- group: the PLA2R-MN patients with negative antibody (Ab); Ab + group: the PLA2R-MN patients with positive anti-PLA2R antibodies. A. The antigen specificity of purified anti-PLA2R antibodies was detected by Western blot with recombinant human PLA2R as antigen, under nondenatured nonreduced condition (a) or denatured reduced condition (b). Strip 1: positive control (plasma from a patient positive of anti-PLA2R antibodies); strips 2–3: purified anti-PLA2R antibodies from the patients in Ab + group; strips 4–7: purified anti-PLA2R antibodies from the patients in Ab- group; strip 8–15: purified anti-PLA2R antibodies from healthy controls; strip 16: negative control (plasma from a healthy donor). The plasma and purified antibodies from Ab + group were diluted at 1:100, and the purified antibodies from Ab- group and HC group were diluted at 1:4. B. The binding of purified anti-PLA2R antibodies to PLA2R on the cell membrane of podocytes was demonstrated by indirect immunofluorescence on human immortalized podocytes. 1. positive control (polyclonal rabbit anti-PLA2R antibodies, 1:200); 2. purified anti-PLA2R antibodies from Ab + group (1:50); 3: purified anti-PLA2R antibodies from Ab- group (1:4); 4–5: purified anti-PLA2R antibodies from HC group (1: 4); 6: negative control (normal human IgG, 1:50)