Videos from Wagner and Glotzer


Video 1: Light-mediated membrane recruitment of optoGEF. 

A NIH3T3 cell expressing Stargazin-GFP-LOVpep and optoGEF was locally photoactivated with 405 nm light every 20 sec with a 960 ms pulse (yellow box). To visualize optoGEF recruitment, 561 nm images were taken every 20 sec before (5 min), during (5 min), and post (5min) photoactivation. 



Video 2: Light-mediated activation of RhoA induces actin polymerization. 

A NIH3T3 cell expressing Stargazin-GFP-LOVpep, optoGEFYFP, and mApple-actin was locally photoactivated with 405 nm light every 20 sec with a 960 ms pulse (yellow box). To visualize the effects of RhoA activation on the actin network, 561 nm images were taken every 20 sec before (15 min), during (15 min), and post (15min) photoactivation. 



Video 3: Light-mediated activation of RhoA induces myosin II accumulation. 

A NIH3T3 cell expressing Stargazin-GFP-LOVpep, optoGEFYFP, and mApple-MLC was locally photoactivated with 405 nm light every 20 sec with a 960 ms pulse (yellow box). To visualize the effects of RhoA activation on myosin localization, 561 nm images were taken every 20 sec before (15 min), during (15 min), and post (15min) photoactivation. 



Video 4: Light-mediated activation of RhoA in non-contractile anaphase HeLa cell induces furrow formation at the midzone. 

A non-contractile anaphase HeLa cell (200 nM BI 2536) expressing Stargazin-GFP-LOVpep and optoGEF was locally photoactivated with 405 nm light every 20 sec with a 960 ms pulse (yellow box) at the midzone. To visualize optoGEF recruitment and furrow induction, 561 nm images were taken every 1 min during (20 min) and post (6 min) photoactivation. 



Video 5: Light-mediated activation of RhoA in non-contractile anaphase HeLa cell induces furrow formation at the poles. 

A non-contractile anaphase HeLa cell (200 nM BI 2536) expressing Stargazin-GFP-LOVpep and optoGEF was locally photoactivated with 405 nm light every 20 sec with a 960 ms pulse (yellow box) at the poles. To visualize optoGEF recruitment and furrow induction, 561 nm images were taken every 20 sec during (23 min) and post (14 min) photoactivation. 



Video 6: Simultaneous photoactivation of midzone and polar regions induces a similar response in non-contractile anaphase HeLa cells. 

A non-contractile anaphase HeLa cell (200 nM BI 2536) expressing Stargazin-GFP-LOVpep and optoGEF was simultaneously photoactivated with 405 nm light every 20 sec with a 960 ms pulse (yellow box) at both the midzone and polar regions. To visualize optoGEF recruitment and furrow induction, 561 nm images were taken every 20 sec during (20 min) and post (5 min) photoactivation. 



Video 7: Exogenous activation of RhoA in the poles competes with endogenous furrow formation. 

A dividing HeLa cell expressing Stargazin-GFP-LOVpep and optoGEF was locally photoactivated with 405 nm light every 20 sec with a 960 ms pulse (yellow box) at the poles. To visualize optoGEF recruitment and the effect on endogenous furrow formation, 561 nm images were taken every 20 sec during (30 min) photoactivation. 



Video 8: Light-mediated activation of RhoA induces furrow formation during metaphase. 

A metaphase HeLa cell expressing Stargazin-GFP-LOVpep and optoGEF was locally photoactivated with 405 nm light every 20 sec with a 960 ms pulse (yellow box). To visualize optoGEF recruitment and furrow induction, 561 nm images were taken every 20 sec during (23 min) and post (14 min) photoactivation. 



Video 9: Light-mediated activation of RhoA induces rapid and nearly complete furrow formation in non-adherent interphase NIH3T3 cells. 

A non-adherent interphase NIH3T3 cell expressing Stargazin-GFP-LOVpep and optoGEF was locally photoactivated with 405 nm light every 20 sec with a 960 ms pulse (yellow box). To visualize optoGEF recruitment and furrow induction, 561 nm images were taken every 1 min during (20 min) and post (10 min) photoactivation. 



Video 10: Low levels of global RhoA activation dampen the extent and rate of furrow induction upon local RhoA activation in non-adherent interphase NIH3T3 cells. 

A non-adherent interphase NIH3T3 cell expressing Stargazin-GFP-LOVpep and optoGEF was photoactivated both globally (10 ms pulse) and locally (960 ms pulse, yellow box) with 405 nm light every 20 sec. To visualize optoGEF recruitment and furrow induction, 561 nm images were taken every 20 sec during (10 min) and post (6 min) photoactivation.



Glotzer Lab | Department of Molecular Genetics and Cell Biology | University of Chicago          © Michael Glotzer 2015