Negative and positive controls for colocomotion analysis. Left: as a negative control, SNAPf-MBP-TMD and Halo-MBP-TMD expressed in HeLa cells were labeled with HTL-TMR (red channel) and SNAPsurfaceDY647 (blue channel), respectively, and imaged with dual-color TIRFM (Olympus IX71; frame rate, 30 Hz; playback, real time). Right: as a positive control, HaloTag-SNAPf-IFNAR2 expressed in HeLa cells was labeled with HTL-TMR (red channel) and SNAPsurfaceDY647 (blue channel) and imaged by dual-color TIRFM. Colocomotion trajectories (≥10 steps) are depicted in yellow.
