Quantum dot ligands provide new insights into erbB/HER receptor−mediated signal transduction

Supplementary Fig. 1 (pdf 27k)
Supplementary Fig. 1

Supplementary Movie 1. EGF-QD binding and internalization (Fig2a). Time series of CHO cells expressing erbB1-eGFP (green) with addition of 200 pM 6:1 EGF-QD (red). EGF-QD binding and internalization of the erbB1-eGFP-EGF-QD complex is observed. Note that the cell in the upper left corner undergoes cell division during EGF-QD-erbB1-eGFP endocytosis. 4 s/frame, playback 10 frames/s. Resolution: 0.14 -m x 0.14 m. (mov 3952k)
Supplementary Movie 1. EGF-QD binding and internalization (Fig2a). Time series of CHO cells expressing erbB1-eGFP (green) with addition of 200 pM 6:1 EGF-QD (red). EGF-QD binding and internalization of the erbB1-eGFP-EGF-QD complex is observed. Note that the cell in the upper left corner undergoes cell division during EGF-QD-erbB1-eGFP endocytosis. 4 s/frame, playback 10 frames/s. Resolution: 0.14 -m x 0.14 m.

Supplementary Movie 2. Z-stack projection of EGF-QD activation and internalization (Fig. 2b). Time series of CHO cells expressing erbB1-eGFP (green) with addition of 250 pM 30:1 EGF-QD (red). Ruffling of the cell surface in response to the EGF-QD binding is seen, followed by extensive internalization. Maximum projection of 12 slices to capture whole cell. 1 min/frame, playback 10 frames/s. _x,y,z: 0.14 m x 0.14 m x 0.62 m. (mov 556k)
Supplementary Movie 2. Z-stack projection of EGF-QD activation and internalization (Fig. 2b). Time series of CHO cells expressing erbB1-eGFP (green) with addition of 250 pM 30:1 EGF-QD (red). Ruffling of the cell surface in response to the EGF-QD binding is seen, followed by extensive internalization. Maximum projection of 12 slices to capture whole cell. 1 min/frame, playback 10 frames/s. _x,y,z: 0.14 m x 0.14 m x 0.62 m.

Supplementary Movie 3. Selective internalization of EGF-QD-erbB1 on erbB3-mCitrine expressing cells (Fig. 3a). Time series of an A431 cell expressing erbB3-mCitrine (green) with addition of 200 pM 6:1 EGF-QD (red). EGF-QDs are seen to bind to the filopodia and travel to the cell body, while the erbB3-mCitrine remains on the cell surface. Maximum projection of 4 slices. 4.5 s/frame, playback 10 frames/s. _x,y,z: 0.1 m x 0.1 m x 0.5 m. (mov 1076k)
Supplementary Movie 3. Selective internalization of EGF-QD-erbB1 on erbB3-mCitrine expressing cells (Fig. 3a). Time series of an A431 cell expressing erbB3-mCitrine (green) with addition of 200 pM 6:1 EGF-QD (red). EGF-QDs are seen to bind to the filopodia and travel to the cell body, while the erbB3-mCitrine remains on the cell surface. Maximum projection of 4 slices. 4.5 s/frame, playback 10 frames/s. _x,y,z: 0.1 m x 0.1 m x 0.5 m.

Supplementary Movie 4. ErbB1-eGFP-EGF-QD endosome fusion (Fig. 4). Time series of a CHO cell expressing erbB1-eGFP (green). Series begins 30 min after addition of 200 pM 6:1 EGF-QD (red). Brownian motion, directed movement and fusion of the erbB1-eGFP-EGF-QD endosomes is observed. Maximum projection of 3 slices. 3.6 s/frame, playback 5 frames/s. _x,y,z: 0.14 m x 0.14 m x 0.5 m. (mov 585k)
Supplementary Movie 4. ErbB1-eGFP-EGF-QD endosome fusion (Fig. 4). Time series of a CHO cell expressing erbB1-eGFP (green). Series begins 30 min after addition of 200 pM 6:1 EGF-QD (red). Brownian motion, directed movement and fusion of the erbB1-eGFP-EGF-QD endosomes is observed. Maximum projection of 3 slices. 3.6 s/frame, playback 5 frames/s. _x,y,z: 0.14 m x 0.14 m x 0.5 m.

Supplementary Methods (pdf 32k)
Supplementary Methods

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