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. 2022 Mar 7;221(4):e202105107. doi: 10.1083/jcb.202105107

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© 2022 Varadarajan et al.

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Figure S2. — Optogenetic stimulation of active RhoA induces whole-field tissue contraction. (A) Whole-field 405-nm laser light stimulation (white dashed box) of an embryo expressing prGEF-mCherry (2xPDZ-mCherry-LARG(DH), gray) and GFP-LOVpep induces whole-field recruitment of prGEF-mCherry to junctions. (B) Quantification of experiments in A. Mean normalized intensity of prGEF-mCherry (2xPDZ-mCherry-LARG(DH)) and GFP-LOVpep upon whole-field 405-nm laser stimulation (gray shaded box, 300–600 s). Shaded regions represent SEM; GFP-LOVpep: n = 9 junctions, 3 embryos, 3 experiments; prGEF-mCherry: n = 12 junctions, 3 embryos, 3 experiments. (C) Time projection images of an embryo expressing active Rho probe (mCherry-2xrGBD, gray). Whole-field light stimulation (white dashed box) leads to increased active Rho at junctions and induces tissue contraction. Zoomed images of cells are indicated by yellow dashed boxes. (D) Quantification of experiments in C. Mean normalized intensity of active Rho probe (mCherry-2xrGBD) upon whole-field 405-nm laser stimulation (gray shaded box). Shaded regions represent SEM; n = 30 junction, 3 embryos, 3 experiments. (E) Site-specific light stimulation of a junction (white dashed box) expressing prGEF-mCherry (2xPDZ-mCherry-LARG(DH), gray) and GFP-LOVpep induces site-specific recruitment of prGEF-mCherry to the junction.