Fig. 5

Long-term depletion of ROS impairs myosin activation and epithelial cell morphology. (A) Workflow of MRLC distribution and cell morphology quantification before amputation and immunofluorescence staining of p-MRLC postamputation in MO-injected groups. (B) Representative morphology of the duox morphant and p53 control groups. Scale bar, 500 μm. (C) Representative images of immunofluorescence of p-MRLC for MO-injected groups unwounded (UW) or at 2 and 15 mpa. Embryos were also stained with phalloidin and DAPI to reveal F-actin and the nucleus. Scale bar, 50 μm. (D and E) F-Actin and p-MRLC intensity of morpholino-injected groups at the indicated time points. Data are presented as mean ± SE, pooled from two independent experiments (p53 control UW group at n = 18, duox + p53 UW group at n = 20, p53 control group at 2 mpa [n = 41], duox + p53 group at 2 mpa [n = 40], p53 control group at 15 mpa [n = 35], duox + p53 group at 15 mpa [n = 34; N = 2]). (F) Representative images of MRLC and plasma membrane in suprabasal epithelia for MO-injected groups. Scale bar, 5 μm. (G and H) Quantification of the suprabasal epithelial cell membrane morphology for MO-injected groups. Data are presented as mean ± SE, pooled from three independent experiments (n = 20; N = 3). (D and E) Two-way ANOVA. (G and H) Unpaired t-test.