Fig. 3: Effects of tropomyosin-1 and tropomyosin-3 depletions on cell morphology, force production, migration, and the actin cytoskeleton.

a Representative wide-field images of wild-type, Tpm1 knockout, and Tpm3 knockout U2OS cells stained for F-actin (phalloidin) illustrating the morphological differences between the Tpm1 knockout and Tpm3 knockout cells. The arrows highlight the abnormal tails of the Tpm3 knockout cells. Scale bars, 20 µm. b Representative wide-field images of wild-type, Tpm1 knockout, and Tpm3 knockout cells stained for F-actin (phalloidin), demonstrating defects in the stress fiber networks of the Tpm1 knockout and Tpm3 knockout cells. Scale bars, 20 µm. c Cell circularity analysis of wild-type (n = 42), Tpm1 knockout; clone 1 (C1) (n = 44) clone 2 (C2) (n = 45), and Tpm3 knockout; clone 2 (C2) (n = 37) clone 11 (C11) (n = 38) cells after 90 min of plating. The data represents mean ± SE. The exact p-values (one-way ANOVA followed by Turkey multiple comparison test): p = 0.058; 0.00000003; 0.0000010. d Representative traction force maps of wild-type, Tpm1 knockout and Tpm3 knockout cells plated on 9.6 kPa hydrogels coated with fibronectin. Scale bar 20 µm. e Mean traction (per cell) of wild-type (n = 43), Tpm1KO (n = 41) and Tpm3KO (n = 42) cells on 9.6 kPa hydrogels coated with fibronectin. The data represents mean ± SE. The exact p-values (two-tailed Student’s t test): p = 0.000015; 0.0000035; 0.14. f Random migration trajectory maps of wild-type, Tpm1 knockout, Tpm3 knockout cells plated on fibronectin-coated substrate. Please, note the differences in the scales of x- and y-axes between wild-type, Tpm1 and Tpm3 panels. g, h Analysis of random migration rates and cell directionality of wild type (n = 30), Tpm1 knockout; clone 1 (C1) (n = 42) clone 2 (C2) (n = 27), and Tpm3 knockout; clone 2 (c2) (n = 55) clone 11 (C11) (n = 34) cells. The data represents mean ± SE. The exact p-values (one-way ANOVA followed by Turkey multiple comparison test) are provided in the Source data.