Supp. Mat. Volume 7 Issue 1 January 2006 - 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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Supplemental Material

Volume 7 issue 1 January 2006
Dynamics of MAL2 During GPI-anchored Protein Transcytotic Transport to the Apical Surface of Hepatoma HepG2 Cells
María C. de Marco, Rosa Puertollano, José A. Martínez-Menárguez and Miguel A. Alonso

SUPLEMENTARY MATERIAL:

Movie 1: Dynamics of MAL2 analysed by fluorescence recovery after photobleaching at the canalicular region. The GFP-MAL2 label at the canalicular membrane (arrow) of HepG2/GFP-MAL2 cells was photobleached. MAL2 was transported from the globular structure to the canalicular area by tubular and vesicular carriers. Still images from this movie are presented in the top panels of Figure 4A. Frames were collected every 3 seconds. The display rate is 5 frame/second.

Movie 1 (.mov)

Movie 2: Dynamics of MAL2 analysed by fluorescence recovery after photobleaching at the MAL2+ globular structure. The GFP-MAL2 label at the MAL2+ globular structure (arrow) of HepG2/GFP-MAL2 cells was photobleached. MAL2 was transported from the canalicular area to the globular structure by tubular and vesicular carriers. Note that one of the cells lacks a globular structure at the beginning of the experiment but assembles one during the recovery of the photobleached globular structure present in the other cell. Still images from this movie are presented in the bottom panels of Figure 4A. Frames were collected every 3 seconds. The display rate is 5 frame/second.

Movie 2 (.mov)

Movie 3: Dynamics of the bile canaliculus of HepG2 cells. The contrast-phase image of a pair of HepG2 cells forming a bile canaliculus (arrow) was recorded at different times. Frames were collected every 20 minutes. The display rate is 2 frame/second.

Movie 3 (.mov)

Movie 4 : Dynamics of MAL2 and CD59 during transcytosis of the latter. Representative experiment in which GFP-MAL2 and CD59 movements were recorded during basolateral-to-apical transcytosis of CD59 in HepG2/GFP-MAL2 cells. Note the redistribution of MAL2 to the periphery, the clustering of internalized CD59 within and around the peripheral structures with MAL2, and the formation of the globular structure containing CD59 and MAL2 and its subsequent movement to the canalicular membrane. Still images from this movie are presented in Fig. 5A,B. Frames were collected every 10 minutes. The display rate is 1 frame/second.

Movie 4 (.mov)