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Fig. 4. Proton uptake and fusion of vacuoles. (A) Proton uptake of wild-type vacuoles (mixture of the fusion tester strains OMY1 and DKY6281) and vacuoles from {Delta}vtc1 (OMY2/OMY5) was measured. (B) Proton uptake and fusion activity of wild-type vacuoles (OMY1/DKY6281 or SBY86/SBY85, respectively) were compared with those of vacuoles from {Delta}vtc1 (OMY2/OMY5), {Delta}vtc2 (OMY4/OMY7), {Delta}vtc3 (OMY3/OMY6) and {Delta}vtc4 (SBY83/SBY82). Proton uptake activity of the wild-type vacuoles was set to 100%. 100% wild-type control fusion was 3.85 U (OMY1/DKY6281) and 3.08 U (SBY86/SBY85), respectively. (C) Comparison of proton uptake activity and fusion activity of wild-type vacuoles (OMY1/DKY6281) in the presence of different concentrations of concanamycin A. Values were plotted as a percentage of the control (vacuoles without concanamycin A). n=3. Control fusions were 2.93 U, 2.40 U and 4.11 U. (D) Proton uptake and fusion activity of wild-type vacuoles (OMY1/DKY6281) with either control antibody or antibodies to Vtc4p. The antibody concentration was 60 µM (c.f. Muller et al., 2002). Proton uptake activity of the sample with control antibodies was set to 100%.