Figure 4
From: CLIMP-63 is a gentamicin-binding protein that is involved in drug-induced cytotoxicity
CLIMP-63 in a cochlear organ of Corti cell line forms the DTT-resistant dimers, and gentamicin induces the dimerization of the CLIMP-63 that is dependent on palmitoylation. (a) In the non-permissive condition (39°C), the organ of Corti HEI-OC1 cell line expressed the DTT-resistant dimer form of CLIMP-63 in western blots. However, there was minimal dimer induction in stria vascularis SV-k1 cells in either condition. (b) CLIMP-63 gene expression levels were not altered in HEI-OC1 cells in the non-permissive condition, confirmed by RT-PCR. (c) After gentamicin treatment at 5 mM for 3 days, both KPT11 and KDT3 expressed higher levels of the DTT-resistant dimers, and higher monomer levels in KPT11 cells in western blots. (d) Cells (293T) transfected with the wild-type CLIMP-63 (p63WT) formed the DTT-resistant dimers, but those with palmitoylation-incompetent mutants C100S and C100A (p63CS and p63CA) showed reduced amounts of the dimers compared with the wild type. Gentamicin treatment increased the dimer level only in cells transfected with the wild-type CLIMP-63. (e) COS-7 transfected with the CLIMP-63 wild type (p63WT) or C100A mutant (p63CA) showed microtubule associated expression. The C100S mutant also showed similar expression pattern (data not shown)
