Inhibition of ROCK activity rescues formation of polarized cysts in the absence of DDR1 signaling. Inhibition of ROCK activity rescues formation of polarized.

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Inhibition of ROCK activity rescues formation of polarized cysts in the absence of DDR1 signaling. Inhibition of ROCK activity rescues formation of polarized cysts in the absence of DDR1 signaling. (A) MDCK cells were cultured in 3D collagen gel for 5 d in the presence or absence of DDR1-IN-1 (1 μM), Y27632 (10 μM), and/or H1152 (4 μM). The cells were fixed and stained for GP135 (green) and F-actin (red). The samples were analyzed by confocal microscopy. (B) The cells were cultured as in (A) in the presence of DDR1-IN-1 (1 μM) and/or blebbistatin (25 μM). Bright-field images were captured by a widefield microscope. Arrows point to the lumens. (C) Population of the cysts having monolayer lumen was counted from each treatment and percentages of the cysts within total structures are shown (grey). Total n number of structures incorporated in this analysis are 460 (control), 432 (DDR1-IN-1), 771 (DDR1-IN-1 + Y27632), 250 (DDR1-IN-1 + H1152), 231 (Y27632), 142 (H1152), and 134 (DDR-IN-1 + blebbistatin). Pia Pernille Søgaard et al. LSA 2019;2:e201800276 © 2019 Søgaard et al.