Figure 2.

Wild-Type and Myosin-18B-Depleted Cells Show Similar Actin-Associated Phenotypes When Cells Are Grown on Soft Substrates

(A) The representative imaging and quantification of ventral- and dorsal-associated focal adhesions in wild-type (n = 19 for vFA, n = 20 for dFA) and myosin-18B knockout (n = 19 for vFA, n = 21 for dFA) cells. vFA (red arrows), ventral associated focal adhesion; dFA (orange arrows), dorsal associated focal adhesion.

(B) Representative images of actin filaments and quantification of the total numbers of thick bundle in wild type (n = 27), myosin-18B knockout (n = 27) and CaMKK2 inhibition (n = 19) cells grown on softer matrix (5 kPa). orange arrows, dorsal stress fiber; blue brackets, transverse arcs.

(C) Representative traction force maps and quantification of cell-exerted strain energy of wild-type (n = 19), myosin-18B knockout (n = 23), and CaMKK2 inhibitor-treated cells (n = 27) grown on softer matrix (5 kPa). Scale bars, 10 μm in (A–C).

(D) Representative examples (corresponding to Video S1, S2, S3, and S4) and quantified centripetal flow rates of transverse arcs in wild-type cells (n = 27), myosin-18B knockout cells (n = 25), full-length (FL) myosin-18B rescue cells (n = 25), and CaMKK2 inhibitor treated cells (n = 25) expressing GFP-actin grown on softer matrix (5 kPa). Yellow arrows, the positions of the observed arcs in the beginning of the videos; red arrows, the positions of the same arcs in subsequent time-lapse images. Scale bar, 5 μm.

Quantification data are represented as mean ± SEM. ∗∗∗p < 0.001 (Mann-Whitney-Wilcoxon rank-sum test).