File:Intermolecular-biparatopic-trapping-of-ErbB2-prevents-compensatory-activation-of-PI3KAKT-via-ncomms11672-s2.ogv

English: Supplementary Movie 1 Induction of Apoptosis by Bi-paratopic DARPins in Comparison to Trastuzumab Treatment. (S1-S2) BT474 cells were seeded 24 h before treatment in RPMI1640 containing 10 % FBS in a 12- well dish. Annexin V-Alexa488 (Life Technologies) was added to give a 1:50 final dilution and propidium iodide (2 g/l) was added to give a 1:2000 final dilution. Afterwards, cells were treated with 100 nM of trastuzumab. Phosphatidylserine flipping (green fluorescence, early apoptosis), loss of cell membrane integrity (red fluorescence, late apoptosis) and bright field images were recorded each 5 min for 48 h on a Lumascope 600 (etaluma) with the lowest possible LED power. Composite images were obtained by the manufacturer's software and videos were put together using the time lapse assembler.