Fig. 5: Overcome the influence of super high density of ligands via Rac1 signaling.
From: Control cell migration by engineering integrin ligand assembly
a F-actin phalloidin (magenta), vinculin N-terminal ___domain lacking the tail ___domain (vin258) (cyan), and paxillin (yellow) immunofluorescence in HuH-7 cell expressing pEFGPC1/GgVcL 1-258 upon the treatment of FFFIKLLI (100 μM) for 12 h. Scale bar represents 5 μm. b Kymograph of normalized edge velocity and mean velocity over time for protrusions and retractions of HuH-7 cells expressing pEGFPC1/GgVcL 1-258 with or without the treatment of FFFIKLLI for 12 h. n = 10 cells. Data are presented as mean ± s.d. c The trajectory plots of ~200 randomly selected migrating HuH-7 cells expressing pEGFPC1/GgVcL 1-258 with or without the treatment of FFIKLLI. Scale bars represent 50 μm. F-actin phalloidin staining (magenta) and paxillin (green) immunofluorescence (three independent experiments were performed) d, kymograph of normalized edge velocity and mean velocity over time for protrusions and retractions (n = 9, 12 cells, respectively, data are presented as mean ± s.d.) e, and the trajectory plots (n = ~200) f of HuH-7 cells pretreated with Rho Activator II (1 μg/mL) with or without 12 h treatment of FFFIKLLI (100 μM). F-actin phalloidin staining (magenta) and paxillin (green) immunofluorescence (three independent experiments were performed) g, kymograph of normalized edge velocity and mean velocity over time for protrusions and retractions (n = 7, 8 cells, respectively, data are presented as mean ± s.d.) h, and the trajectory plots (n = ~200) i of HuH-7 cells with elevated Rac1 activation with or without 12 h treatment of FFFIKLLI (100 μM). j Schematic summary of outside-in regulation of HuH-7 cell motility via precise control of ligand density on nanofilaments, and the restoration of cell motility via intracellular signaling. Source numerical data are available in source data.
