Supp. Mat. Volume 7 Issue 4 April 2006 (b) - Traffic the International Journal of Intracellular Transport

Notch receptors are clustered and trans-endocytosed by Delta ligand cells. Confocal micrograph of a Delta expressing cell (left) interacting with a Notch expressing cell (right). Following interaction with Delta (blue), cell surface Notch (yellow) is clustered at cell-cell interfaces. Notch extracellular domain is detected within Delta cells (green) indicative of trans-endocytosis. Endocytosis of ligand while bound to Notch may produce a force sufficient to pull Notch apart and activate signaling.

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Volume 7 issue 4 April 2006
A multigene family encoding R-SNAREs in the ciliate Paramecium tetraurelia
Christina Schilde, Thomas Wassmer, Joerg Mansfeld , Helmut Plattner, and Roland Kissmeh

SUPLEMENTARY MATERIAL:

Supplemental Figure 1:
Synaptobrevin family phylogeny based on only the SNARE domain. Neighbour joining tree with 1000 bootstrap replicates encompassing the SNARE domains of P. tetraurelia synaptobrevin proteins (Pt Sybs; see Table 1), T. thermophila (Tt ) synaptobrevin homologs from preliminary gene predictions 08-2004 at TIGR (gene identifiers 2415, 4410, 4411, 4722, 7285, 11170, 14913, 19728, 20867), Arabidopsis thaliana (At) VAMPs 711-714, 723-725, 727 (GenBank accession numbers O49377, Q9SIQ9, Q9LFP1 , Q9FMR5, Q8VY69, O23429, O48850, Q9M376), Homo sapiens (Hs) synaptobrevin 1 (Syb1), VAMP2, VAMP3, Ykt6 and synaptobrevin-like protein 1 (Sybl1) (GenBank accession numbers AAA60603, AAH02737, CAB63146, CAG46805, AAH56141), Saccharomyces cerevisiae (Sc) Snc1, Snc2, Nyv1, Ykt6 and Sec22 (GenBank accession numbers AAC05002, NP_014972, NP_013194, AAB32050, AAB6737), Dictyostelium discoideum (Dd ) SybA, VAMP7A, VAMP7B, ( DDB0214903, DDB0231535, DDB0231542) and DDB0190688, DDB0219546, and DDB0219666 and Entamoeba histolytica (Eh) synaptobrevin-like proteins 1 and 2 (Sybl1 and Sybl2) (GenBank accession numbers AY256852 and AY309014). Paramecium synaptobrevins addressed in this work are indicated with grey diamonds. Boostrap support values are indicated above the branches and an evolutionary distance scale is given below the figure.

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Movie 1: Movie file showing the phenotype and swimming behaviour of PtSyb2 gene silenced cells. One mock-silenced cell was added for comparison. PtSyb2 silenced cells at 48 hours of silencing have a rounded shape and perform quick rotations around their own axis before they eventually die.

Movie 1 (.avi)

Supplemental Table 1: Oligonucleotide primers used for amplification and cloning of P. tetraurelia synaptobrevin genes. Non-homologous bases are indicated in italics, added restriction sites are shown in bold.

View Table 1 (.doc)