Fig. 1: Development of the OptoMYPT system. | Nature Communications

Fig. 1: Development of the OptoMYPT system.

From: Optogenetic relaxation of actomyosin contractility uncovers mechanistic roles of cortical tension during cytokinesis

Fig. 1

a Schematic illustration of the signaling pathway for the regulation of MLC phosphorylation. b Domain structure of human MYPT1. c Schematic illustration of the OptoMYPT system. Stargazin-mEGFP-iLID is anchored to the plasma membrane. Upon blue light illumination, the SspB-mScarlet-I-fused PP1c-binding domain (PP1BD) of MYPT1 translocates to the plasma membrane along with endogenous PP1c, and inactivates NMII at the cell cortex. d Representative images of the SspB-mScarlet-I or the indicated SspB-mScarlet-I-PP1BDs of MYPT1 (upper two rows) and simultaneously expressing PP1c-miRFP703 (lower two rows) in MDCK cells under the dark condition (the first and third rows) and blue light condition (the second and fourth rows). Stargazin-mEGFP-iLID was also expressed as a localizer in all experiments. The yellow arrowhead indicates a cell showing nuclear accumulation of SspB-mScarlet-I-MYPT38. Blue light was continuously illuminated from the top of the stage. Scale bar, 20 μm. e Quantification of the cytoplasmic fluorescence change in mScarlet-I in panel d. The average values (bold lines) are plotted as a function of time with the SD. n = 15 cells. f Quantification of the cytoplasmic fluorescence change in PP1c-miRFP703 in the indicated MDCK cells. The average values (bold lines) are plotted as a function of time with the SD. n = 15 cells.

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