Western blot and autoradiograph of radiolabelled chimeric human/rat α3(IV)NC1 domains. COS supernatants from transfected cells were separated on 12.5% SDS–PAGE gels, which were then electroblotted to nitrocellulose or dried for exposure to autoradiograph film. (a) Western blot showing binding of Goodpasture autoantibodies to radiolabelled chimeric human/rat α3(IV)NC1 domains. (b) Autoradiograph showing the expression levels of chimeric human/rat α3(IV)NC1 domains. The radiolabelled recombinant protein in each lane was: lane 1, rat α3(IV)NC1 (α333); lane 2, αRRH; lane 3, αRHR; lane 4, αHRR; lane 5, αHHR; lane 6, αHRH; lane 7, αRHH; lane 8, human α3(IV)NC1; lane 9, negative control.