Extended Data Fig. 1: Characterization of ATV+ and ATV- in live cells.
From: Optogenetic manipulation of cellular communication using engineered myosin motors
a, Representative images of randomly chosen mouse fibroblasts expressing either (upper) ATV+ or (lower) ATV- with pmBFP chosen from 3 independent experiments. Scale, 20 µm. b, c, Mouse fibroblasts co-expressing sfGFP-ATV+, mKate2-ATV- and pmiRFP in the same cells. ATV+ and ATV- display significantly different localization patterns within the same cells. Representative images chosen from 3 independent experiments. Scale, 20 µm. Mouse fibroblasts expressing (d) sfGFP-ATV+ or (e) sfGFP-optoATV- were imaged under a total internal reflection (TIRF) microscope. d, Left: A kymograph of a cellular protrusion showing single molecule traces of sfGFP-ATV+ moving down the tip. Scale, 1 µm. Right: Quantification of the velocity of ATV+ moving within cellular protrusions. Single dots represent single molecule traces chosen from 3 independent experiments (N = 23 traces). e, Left: A kymograph of a cellular protrusion where single molecule traces of sfGFP-optoATV- retracting back to the cell body upon light stimulation. Scale, 100 nm. Right: Quantification of the velocity of activated optoATV- moving within cellular protrusions. Single dots represent single molecule traces chosen from 3 independent experiments (N = 16 traces). The graphs are shown as mean ± SEM.
