Localization of different Rab GTPases with amino acid substitutions in the N(T)KxD nucleotide binding motif. (A & B) NIH 3T3 cells were transfected with pCMV5 vectors encoding different myc-tagged Rab proteins with the indicated amino acid substitutions. Twenty-four hours after transfection, cells were processed for immunofluorescence microscopy, using the anti-myc monoclonal antibody, followed by FITC-conjugated GAM IgG. Non-transfected cells cannot be seen at the exposure setting used for the photographs. The bar in the upper left panel is 10 microns. (C) Autoradiogram of a thin-layer plate showing 32P-labeled GDP and GTP eluted from the indicated immunoprecipitated proteins. The immunoblot in the lower panel compares the relative amount of mycRab24 collected in 1/10 of each immunoprecipitate.
Maltese et al. BMC Cell Biology 2002 3:25 doi:10.1186/1471-2121-3-25