Figure 8.
Glomerular eluates from MN patients contain antibodies against podocyte antigens. Podocyte extracts were separated by electrophoresis in reducing (a through e) and nonreducing (f through h) conditions and were studied for reactivity with eluates from microdissected glomeruli. Thirty micrograms of protein was used for the analysis: Part was analyzed with silver stain (a and f), and part was incubated with glomerular eluates from patients with MN and then developed with anti-IgG4 (b and g). Eight antigens were recognized in reducing conditions and characterized by mass spectrometry as AR, SOD, and vimentin. Three more bands with a molecular weight of 200, 170, and 155 kD were detected in nonreducing conditions. (c and h) Control lines for Western blot in which glomerular eluates were omitted. (d) Western blot with anti-AR antibody. (e) Western blot with SOD2 antibodies.