Fig. 4: Cph1-PM-dependent changes in collective cell dynamics are mediated by PLD2 activity and membrane tension.
From: Intercellular adhesion boots collective cell migration through elevated membrane tension
a Phosphatidic acid levels for parental MDA and Cph1-PM-MDA cells under red and far-red light in the absence of or on addition of propranolol (Pro) or VU-0285655-1 (VU; all data n = 4 from 4 biological replicates). b Phospholipase D2 (PLD2) hydrolyzes phosphatidylcholine (PC) into phosphatidic acid (PA) and choline and phosphatidic acid phosphatase (PAP) hydrolyze PA to diacylglycerol (DAG). PAP is inhibited by Pro and PLD2 is inhibited by VU. c, d Representative phase-contrast micrographs (top) and velocity vector maps (bottom) for Cph1-PM-MDA cells under red and far-red light after treatment with Pro (c) and VU (d). On the vector map, yellow indicated the maximum and blue the minimum. e Wound closure rate from parental MDA or Cph1-PM-MDA cells under red and far-red light. For each condition, cells are shown before treatment or after treatment with Pro or VU (all data n = 4 from 4 biological replicates). f Correlation length of Cph1-PM-MDA cells without treatment under red (n = 732) and far-red (n = 1553) light, or after treatment with Pro under red (n = 252) and far-red (n = 1588) light, or after treatment with VU under red (n = 2213) and far-red (n = 1209) light from 4 biological replicates. g Oleic acid (OA) incorporates into the lipid bilayer of cell membranes and increases membrane tension. h Phase-contrast micrographs (top) and velocity vector maps (bottom) for Cph1-PM-MDA cells under red and far-red light after treatment with OA. On the vector map, yellow indicated the maximum and blue the minimum. i Wound closure rate from parental MDA cells or Cph1-PM-MDA under red and far-red light without treatment or after treatment with OA (all data n = 4 biological replicates). j Correlation length denotes the persistence length of velocity vector under red and far-red light before and after treatment with OA (no treatment, nred = 732, nfar-red = 1553; OA, nred = 846, nfar-red = 1581; 4 biological replicates). All statistics were analyzed by one-way ANOVA with Bonferroni’s multiple comparisons test. Bar plots (a, e, i) are denoted as mean with standard deviation. Box plots (f, j) present the median and 25th and 75th percentiles, and the lower and upper boundaries of whiskers show the 5th and 95th percentiles. Scale bars (c, d, h), 100 µm. Data source and statistical details are provided as a source data file.
