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. 2001 Nov;12(11):3375–3385. doi: 10.1091/mbc.12.11.3375

Figure 3.

Figure 3

Decreased fusion of Vam3ΔN vacuoles. (A) Fusion activity of Vam3ΔN- compared with wild-type-vacuoles. Standard fusion reactions with cytosol and ATP were incubated for 90 min at 26°C, and alkaline phosphatase activity was determined as described in MATERIALS AND METHODS. To allow comparison of independent experiments, wild-type vacuole fusion was set to 100%. Fusion of Vam3ΔN vacuoles was reduced to 40 ± 11% (mean ± SEM, n = 14). (B) Time course of the fusion reaction. Standard fusion reactions (150 μl) containing wild-type or Vam3ΔN tester vacuoles were started with cytosol and ATP. At the indicated time points a 30-μl aliquot was removed and placed on ice to stop the reaction. Fusion activity was measured after 90 min. A representative experiment is shown. (C) Loss of fusion competence over time. Wild-type or mutant BJ3505 and DKY6281 vacuoles were separately incubated with cytosol and ATP at 26°C. At the indicated time points 20 μl of BJ3505 and DKY6281 vacuoles were mixed and incubated for an additional 70 min. Then fusion activity was assayed.