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Probing and manipulating intracellular membrane traffic by microinjection of artificial vesicles

Supporting Information

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  • Download Movie_S01 (MP4) - The movement of injected 20% PS/PC LUVs in a HeLa cell 5 min after injection. HeLa cells were injected with 20% PS/PC LUVs labeled with rhodamine-PE, followed by time-lapse confocal imaging 5 min after microinjection. Time interval between acquired frames was 0.2 s and the acquisition length was 40 s. (Scale bar, 3 µm.)
  • Download Movie_S02 (MP4) - The movement of endogenous endosomes in a HeLa cell. HeLa cells were incubated with Alexa 488-Tfn for 5 min to allow for internalization. Thereafter the cells were washed, followed by time-lapse confocal imaging. Time interval between acquired frames was 0.2 s and the acquisition length was 40 s. (Scale bar, 3 µm.)
  • Download Movie_S03 (MP4) - The movement of injected 20% PS/PC LUVs in a HeLa cell 15 min after injection. HeLa cells were injected with 20% PS/PC LUVs, followed by time-lapse confocal imaging 15 min after microinjection. Time interval between acquired frames was 0.2 s and the acquisition length was 40 s. (Scale bar, 3 µm.)
  • Download Movie_S04 (MP4) - The movement of injected 20% PS/PC LUVs in a HeLa cell 60 min after injection. HeLa cells were injected with 20% PS/PC LUVs, followed by time-lapse confocal imaging 60 min after microinjection. Time interval between acquired frames was 0.2 s and the acquisition length was 40 s. (Scale bar, 3 µm.)
  • Download Movie_S05 (MP4) - The movement of endogenous endosomes in a HeLa cell after nocodazole treatment. HeLa cells were treated with 1 µM nocodazole for 30 min and then incubated with Alexa 488-Tfn for 5 min to label early endosomes. After internalization, the cells were washed with PBS, followed by time-lapse confocal imaging. Time interval between acquired frames was 0.2 s and the acquisition length was 40 s. (Scale bar, 3 µm.)
  • Download Movie_S06 (MP4) - The movement of injected 20% PS/PC LUVs in a nocodazole-treated HeLa cell 60 min after injection. HeLa cells were treated with 1 µM nocodazole for 30 min and then injected with 20% PS/PC LUVs. After further incubation for 60 min with 1 µM nocodazole, time-lapse imaging was collected. Time interval between acquired frames was 0.2 s and the acquisition length was 40 s. (Scale bar, 3 µm.)
  • Download Movie_S07 (MP4) - Approach between an endogenous endosome and an injected 4 EE-SNARE proteoliposome. Time-lapse confocal imaging of a HeLa cell shortly after injecting 4-EE-SNARE LUVs labeled with rhodamine-PE (red). To label endogenous early/recycling endosomes, Alexa 488-Tfn (green) was internalized during microinjection for 5 min. Images were collected a few minutes after the microinjection. Time interval between acquired frames was 0.4 s and the acquisition length was 9 s. (Scale bar, 3 µm.)
  • Download Movie_S08 (MP4) - Fusion between an endogenous endosome and an injected 4 EE-SNARE proteoliposome. Time-lapse confocal imaging of a HeLa cell shortly after injecting 4-EE-SNARE LUVs containing 50 mM calcein (red). To label endogenous early/recycling endosomes, Alexa 633-Tfn (blue) was internalized during microinjection for 5 min. Green indicates dequenched calcein signal generated by dilution during fusion. Images were collected for a few minutes after the microinjection. The time interval between the acquired frames was 1 s and the acquisition length was 30 s. (Scale bar, 3 µm.)

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