Figure 5
Figure 5. The pattern and regulation of tractions in neutrophils migrating on a stiffer substrate. (A) PSD plots of tractions at the leading edge (left panel) and the trailing edge (right panel) of a migratory primary neutrophil. PSD plots were generated based on the results from Fourier analysis of the traction values. The y-axis represents the PSD normalized to the highest peak value (1). The x-axis shows the oscillation frequency (Hertz; top) or period (seconds; bottom). Primary cells were allowed to migrate toward chemoattractant-containing micropipette (fMLP, 10μM) on a FN-coated polyacrylamide gel (100 kPa) for 4 to 5 minutes. Six cells were analyzed, and a representative cell is shown. (B) Left panel: Cross-correlation between tractions at the leading edge and the trailing edge against time offset during migration for individual primary neutrophils. Dotted lines indicate zero offset. Data from 3 representative cells are shown. Time bar represents 24 seconds. Right panel: Summary of time offsets between leading edges and trailing edges (n = 6 cells) in primary cells. (C) PSD plots of tractions at the leading edge (left panel) and the trailing edge (right panel) of a migratory primary neutrophil pretreated with Y-27632 (30μM, 30 minutes). PSD plots were generated based on the results from Fourier analysis of the traction values. The y-axis represents the PSD normalized to the highest peak value (1). The x-axis shows the oscillation frequency (Hertz; top) or period (seconds; bottom). Cells were allowed to migrate toward chemoattractant-containing micropipette (fMLP, 10μM) on a FN-coated polyacrylamide gel (100 kPa) for 4 to 5 minutes. Six cells were analyzed, and a representative cell is shown. (D) Left panel: Cross-correlation between tractions at the leading edge and the trailing edge against time offset during migration for individual Y-27632–treated primary neutrophils. Dotted lines indicate zero offset. Data from 3 representative cells are shown. Time bar represents 24 seconds. Right panel: Summary of time offsets between leading edges and trailing edges (n = 6 cells) in Y-27632–treated primary cells.

The pattern and regulation of tractions in neutrophils migrating on a stiffer substrate. (A) PSD plots of tractions at the leading edge (left panel) and the trailing edge (right panel) of a migratory primary neutrophil. PSD plots were generated based on the results from Fourier analysis of the traction values. The y-axis represents the PSD normalized to the highest peak value (1). The x-axis shows the oscillation frequency (Hertz; top) or period (seconds; bottom). Primary cells were allowed to migrate toward chemoattractant-containing micropipette (fMLP, 10μM) on a FN-coated polyacrylamide gel (100 kPa) for 4 to 5 minutes. Six cells were analyzed, and a representative cell is shown. (B) Left panel: Cross-correlation between tractions at the leading edge and the trailing edge against time offset during migration for individual primary neutrophils. Dotted lines indicate zero offset. Data from 3 representative cells are shown. Time bar represents 24 seconds. Right panel: Summary of time offsets between leading edges and trailing edges (n = 6 cells) in primary cells. (C) PSD plots of tractions at the leading edge (left panel) and the trailing edge (right panel) of a migratory primary neutrophil pretreated with Y-27632 (30μM, 30 minutes). PSD plots were generated based on the results from Fourier analysis of the traction values. The y-axis represents the PSD normalized to the highest peak value (1). The x-axis shows the oscillation frequency (Hertz; top) or period (seconds; bottom). Cells were allowed to migrate toward chemoattractant-containing micropipette (fMLP, 10μM) on a FN-coated polyacrylamide gel (100 kPa) for 4 to 5 minutes. Six cells were analyzed, and a representative cell is shown. (D) Left panel: Cross-correlation between tractions at the leading edge and the trailing edge against time offset during migration for individual Y-27632–treated primary neutrophils. Dotted lines indicate zero offset. Data from 3 representative cells are shown. Time bar represents 24 seconds. Right panel: Summary of time offsets between leading edges and trailing edges (n = 6 cells) in Y-27632–treated primary cells.

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