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Traces shown in figures are the average of 2−4 acquisitions using the same experimental solutions, except Figures 7 and 8, which are single traces.
The parental T-REx™ line was used in all experiments involving HeLa cells except Figures 6 and 7, and was cultured in Eagle's minimal essential medium, 10% (v/v) FBS and 5 μg/ml blasticidin, but without tetracycline.
10.7554/eLife.00592.003 Box 1. Definitions v-vesicle : proteoliposome with reconstituted synaptobrevin-2 in all experiments except Figures 1, 2, and 4D where both synaptobrevin-2 and synaptotagmin-1 were reconstituted.
DOI: http://dx.doi.org/10.7554/eLife.00592.003 v-vesicle: proteoliposome with reconstituted synaptobrevin-2 in all experiments except Figures 1, 2, and 4D where both synaptobrevin-2 and synaptotagmin-1 were reconstituted.
High molecular weight RNA was prepared using the hot phenol method for all experiments except Figures 7B, 3B, 3F and Figure 3 figure supplements 1,2 where guanidine thiocyanate (GTC -phenol preparations were used.
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EL performed all experiments (with the exception of immunostaining), analyzed data, contributed all figures and tables except figure 4, and drafted the first manuscript.
All experiments except Figure 4G were conducted at ∼33°C.
Each figure shows one representative dataset for each assay except Figure 1B, Figure 4 and Figure S2, which present compilations of multiple datasets.
All of the climate maps in this article and in the Supplementary material were created directly from using the Climate Wizard (except Figure 14, and Figure S5 which are analytic derivations of Climate Wizard products).
5) Most of the data (except Figure 5b and Figure 6) only show one time point (fluorescence at ten minutes).
All trees were displayed using NJ plot [ 40] except Figure 1 which was viewed using TreeView [ 41].
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CEO of Professional Science Editing for Scientists @ prosciediting.com