The Olympus FluoView FV1000 confocal microscope was equipped with a temperature-controlled CO2 incubation chamber at 37°C and a 60×/1.35 numerical aperture (NA) Oil UPLSAPO objective (Olympus). EphA2-mCitrine and EGFR-mCherry were excited using a 488-nm argon laser (GLG 3135, Showa Optronics) and a 561-nm diode-pumped solid-state (DPSS) laser (85-YCA-020-230, Melles Griot), respectively. Detection of fluorescence emission was restricted with an acousto-optical beam splitter (AOBS) for mCitrine at 498 to 551 nm and for mCherry at 575 to 675 nm. In all cases, scanning was performed in frame-by-frame sequential mode with 2× frame averaging. The pinhole was set to 250 μm.

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