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Figure 6


Fig. 6. Rescue of function by re-expression of Syt I or Syt IX. (A) Immunoblot analysis of wild-type cells, cells that lack Syt I and Syt IX (Syt I/IX-null) and Syt I/IX-null cells that express the pcDNA3-Syt I or pcDNA3-Syt IX rescue vector. (B) Wild-type and Syt I/IX-null cells were transfected with a plasmid expressing ANF-EGFP or Syt I/IX-null cells were co-transfected with a plasmid expressing ANF-EGFP and the pcDNA3-Syt I or pcDNA3-Syt IX rescue vector. Cells were stimulated in depolarization medium and images were acquired at 0.25-second intervals. The data were analyzed as in Fig. 4D and plotted as the sum of the events for ten wild-type, ten Syt I/IX-null, ten Syt I/IX-null + Syt I and ten Syt I/IX-null + Syt IX cells. Re-expression of either Syt I or Syt IX restored Ca2+-dependent DCV exocytosis in the Syt I/IX-null cells. (C) The sum of the average number of fusion events per cell over time (mean ± s.d.) for wild-type (n=40), Syt I/IX-null (n=10), Syt I/IX-null + Syt I (n=20) and Syt IX-null + Syt IX (n=12). {tau} values for fusion in wild-type (31.2±9.1 seconds) and Syt I/IX-null + Syt I (15.6±1.9 seconds) cells were significantly different (P<0.001).





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