Fig. 1. Tension sensor differentially localized to distinct F-actin structures.

a Schematic of the tension sensor (TS), connected to actin filaments, and b control tension sensor (CTS), connected to one actin filament. c TS expressing U2OS cell on a circular micropattern of 0.2 mg/mL gelatin, showing phalloidin-stained actin, GFP (TS), and paxillin. Scale bar is 25 μm. d CTS expressing U2OS cell on a circular micropattern of 0.2 mg/mL gelatin, showing phalloidin-stained actin, GFP (CTS), and paxillin. Scale bar is 25 μm. e Actin and TS GFP of U2OS cell on a circular pattern. Red dash line indicates a stress fiber. Enrichment colormap showing the normalized ratio of TS to actin. Scale bar is 10 μm. f Actin and TS GFP of U2OS cell treated with 50 μM SMIFH2, on a circular pattern. Enrichment colormap showing the normalized ratio of TS to actin. Scale bar is 10 μm. g Radially averaged sensor enrichment (ρ) for TS and TS+SMIFH2 cells (n = 10 for TS cells, n = 13 for TS +SMIFH2 treated cells). Dashed line indicates central-peripheral border. Data are presentd as mean values ±SD. h Sensor enrichment (ρ) in the central, peripheral and SF regions (For central and peripheral results, n = 10 for TS cells, n = 7 for CTS cells, and n = 5 for TS+SMIFH2 cells. For stress fibers, n = 26 for TS, and n = 14 for CTS). Two-way ANOVA was used for significance. (p(TS central and TS + SMIFH2 central) = 0.006, p(TS central and CTS central) = 0.999, p(TS peripheral and TS + SMIFH2 peripheral) = 0.006, p(TS peripheral and CTS peripheral) = 0.999, p(TS + SMIFH2 peripheral and CTS peripheral) = 0.003, p(TS stress fibers and CTS stress fibers) = 0.998. i iPearson colocalization coefficient for actin/RFP(TS) (n = 9) and actin/RFP(CTS) (n = 9) cells. Two-sided t-test between the two samples p(TS and CTS) = 0.2184. One sample two-sided t-test for each column p < 0.0001. Experiments were repeated three times independently. Data are presented as mean values ±SD. Source data are provided as a Source Data file.