Cells were seeded into p35 dishes and simultaneously transfected with 5 μl of 10 μM duplex siRNAs (sequences given in table S1) (TriFECTa Kit DsiRNA Duplex, Integrated DNA Technologies) using Opti-MEM (Invitrogen) and Lipofectamine RNAiMAX (Invitrogen), according to the manufacturer’s instructions. The following day, cells were split and seeded into two glass-bottom microscopy dishes in the medium indicated above for live cell imaging experiments and transfected again with siRNA using the same protocol. After 8 hours, around 30% of the growth medium was exchanged, and the cells were transfected with the plasmids encoding fluorescent proteins as indicated above.

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