Hiroyasu Tsukaguchi, Akulapalli Sudhakar, Tu Cam Le, Trang Nguyen, Jun Yao, Joshua A. Schwimmer, Asher D. Schachter, Esteban Poch, Patricia F. Abreu, Gerald B. Appel, Aparecido B. Pereira, Raghu Kalluri, Martin R. Pollak
Nephrin binding assay. (a) The bottom autoradiograph shows in vitro–translated and labeled mutant and wild-type (WT) podocin after incubation with purified nephrin, immunoprecipitation with an anti-nephrin antibody, transfer to nitrocellulose, and exposure to radiographic film (as described in detail in Methods). Shown above is the result of Western analysis of the same blots using the anti-nephrin antibody for immunodetection. (b) Densitometry of the R229Q and R291W bands from repeated paired experiments. Intensity (with error bars indicating SEM) is given as a percentage of wild-type intensity, which is set by definition at 100%.