Fig. 4: μkiss labeling to visualize time-dependant TfR-clathrin membrane dynamics. | Nature Methods

Fig. 4: μkiss labeling to visualize time-dependant TfR-clathrin membrane dynamics.

From: A paintbrush for delivery of nanoparticles and molecules to live cells with precise spatiotemporal control

Fig. 4

a, Fluorescence image of HaloTag-OregonGreen-labeled clathrin. The arrowhead and dashed outline mark the labeling region. Scale bar 10 μm. b, Close-up view of the region in a postlabeling, showing clathrin (black) and Tf (orange). The result is indicative of behavior from a series of size n = 13. Scale bar 5 μm. c, Profile of fluorescence signal along the path αβγ marked in b, before Tf labeling (above) and after (below). d, Images of one Tf-labeling cycle on an immobile clathrin cluster (upper) and integrated signal of Tf fluorescence for three labeling cycles (bottom). e, Trajectory and time-schematic of a mobile clathrin spot during flight before (blue) and after (gray) Tf labeling. The trajectory for the Tf-labeled TfR is highlighted in orange. An asterisk marks the point of labeling. Scale bars 1 μm. The inset shows the tracked clathrin cluster with colocalized Tf signal. Scale bars 1 μm. Lower axis marks the temporal ascension of time. f, Number and size distribution of Tf-labeled TfR clusters as a function of time, following initial TfR labeling.

Back to article page