DDR1 inhibition increases ROCK activity in cells seeded on glass.

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DDR1 inhibition increases ROCK activity in cells seeded on glass. DDR1 inhibition increases ROCK activity in cells seeded on glass. (A) MDCK cells were seeded at 40% confluency on glass coverslips in the presence or absence of DDR1-IN-1 (1 μM) and/or Y27632 (10 μM) as indicated for 36 h. The cells were stained for ppMLC (Thr18/Ser19) (green) and F-actin (red). The samples were analyzed by widefield microscope with 20× objective lens. Arrows point to the common cortical actin bands in control cells and ppMLC-positive stress fibres in DDR1-IN-1–treated cells. Images were analyzed for the percentage of cells with ppMLC-positive actin bundles along cell–cell contacts. Analysis of 48–78 cells per condition from two independent experiments is shown. (B) MDCK cells stably expressing DDR1 were cultured at 100% confluence in collagen I–coated (1.5 mg/ml) culture inserts for 36 h. The cells were stained for F-actin (white), DDR1 (cytoplasmic domain, red), and ZO-1 (green). The samples were analyzed by confocal microscopy and representative XY and XZ planes shown. (A) apical, (B) basal. Note that DDR1 colocalizes with ZO-1. Pia Pernille Søgaard et al. LSA 2019;2:e201800276 © 2019 Søgaard et al.