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Fig. 3. Inducible overexpression of EGFP:rab11 chimeras. EcR:EGFP-rab11 cells were grown for 72 hours in the presence of 0.125% vehicle (EtOH) or 5 µM ponasterone A (Pon A), then lysed and analysed by immunoblotting. Protein expression was determined using anti-rab11 antibody and ECL and then analyzed by densitometry. The upper bands (~50 kDa) are EGFP-rab11 and the lower bands (~25 kDa) are endogenous rab11. The levels of overexpression of EGFP-rab11WT and the S25N mutant were approximately fourfold and twofold, respectively, that of endogenous rab11. When observed by fluorescence microscopy, more than 80% of cells expressed EGFP-rab11 and their subcellular distributions were similar to those observed in transiently transfected cells (data not shown).





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