Figure 1
From: CLIMP-63 is a gentamicin-binding protein that is involved in drug-induced cytotoxicity
Gentamicin-binding protein identification. (a) Proximal tubule KPT11 cells and distal tubule KDT3 cells were compared for cell viability after gentamicin treatment for 3 days by MTT assay. KPT11 cells had significantly lower viability compared with KDT3 cells following 10 mM gentamicin treatment (**P<0.01). (b) Gentamicin (10 mM) induced significantly more caspase-3 activity in KPT11 cells compared with KDT3 cells (*P<0.05). Data in a and b are shown as mean±S.D. of three independent experiments. (c) GBPs were pulled down from total cell lysates of KPT11 and KDT3 cells. SDS-polyacrylamide gel electrophoresis followed by Coomassie blue staining revealed several GBP bands specific to KPT11 cells (representative gel from three experiments). Mass spectrometric analysis of peptides from excised gel bands identified these GBPs with the highest numbers of unique peptides. (d) Western blotting on the pull-down samples confirmed the GBPs identified by mass spectrometry
