Figure 3
From: CLIMP-63 is a gentamicin-binding protein that is involved in drug-induced cytotoxicity
Different expression levels of CLIMP-63, oligomerization of CLIMP-63, and gentamicin binding to the CLIMP-63 lumenal domain. (a) RT-PCR of specified mouse tissues revealed varying CLIMP-63 expression levels. (b) RT-PCR of mouse kidney cell lines show higher levels of CLIMP-63 mRNA expression in proximal tubule cells compared with distal tubule cells. (c, left panel) Proximal, but not distal, tubule cells express DTT-resistant dimers of CLIMP-63, revealed by western blotting. (c, right panel) Without DTT, all samples displayed bands that correspond to dimers and tetramers. (d) The GST fusions of the cytosolic domain (p63-Cyto) and lumenal domain (p63-lumen) had approximate molecular masses of 42 and 75 kDa, respectively, as revealed by Coomassie blue staining (shown as ‘input’). p63-lumen was efficiently pulled down, whereas negligible GST or weak p63-Cyto proteins were pulled down by gentamicin–agarose (GT). There were no GST fusions pulled down by agarose without gentamicin conjugation (no GT), although a weak GST band was detected
