FIGURE

Fig. 1

ID

ZDB-FIG-220906-12

Publication

Toh et al., 2022 - Optogenetic control of YAP cellular localisation and function

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Fig. 1

A. Schematic of optoYAP construct. OptoYAP is folded in the dark due to the interaction between LOV2 domain and Jα helix. Under 488 nm light, nuclear localisation signal (NLS) is exposed and optoYAP is transported into the nucleus. Activation protocol was performed with 1 s blue light pulse every 30 s (blue bars), and cells were imaged every minute (red bars) for 20 min.
B. Representative images of mCherry-optoYAP in HEK293T cells imaged with only 561 nm laser following the protocol in (A). Scale bars, 10 μm.
B′. Fold-change in nuclear localisation of mCherry-optoYAP (n = 50 cells from three independent experiments).
C. Representative images of the same cells in (B) exposed to both 488 and 561 nm following the protocol shown in (A). Scale bars, 10 μm.
C′. Fold-change in nuclear localisation of the mCherry-optoYAP (n = 50 cells from three independent experiments).
D. HEK293T cells transfected with optoYAP were subjected to three cycles of activation protocol and recovery in the dark. (Top) The ratio of mCherry-optoYAP signal in the nucleus to the cytoplasmic mCherry-optoYAP signal (n = 8 cells from two independent experiments). (Bottom) Representative images of the mCherry-optoYAP signal from the same HEK293T cells at each cycle. Scale bars, 10 μm.
E. Activation time constant (τ_a) and recovery time constant (τ_r) (see Materials and Methods and Fig EV1B and C) of optoYAP in HEK293T cells. Vertical dashed line represents time when 488 nm stimulation ceased. Red line indicates average nuclear/cytoplasmic ratio (n = 12 cells).
F. HEK293T cells transfected with optoYAP subjected to 0.065, 0.163 and 0.325 mW 488 nm laser power at the focal plane. N = 3 cells per laser power.

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Phenotype Detail

Acknowledgments

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