Figure 6
Palladin knockdown cells show more efficient recovery from Blebbistatin treatment.
(A) DIC image of a palladin KD (Palld4) cell on a gel of intermediate stiffness (10–30 kPa range). Scale bar: 10 μm. (B) DIC image of the same cell as in A, 30 minutes after incubation in 15 μM blebbistatin. (C) DIC image of the cell 1 hour after washout from blebbistatin, showing recovery of cell morphology. (D) Traction force map of the cell in A showing robust generation of traction forces. (E) Traction force map of the cell in B, showing disappearance of traction forces upon blebbistatin addition. (F) Traction force map of the cell in C, showing recovery of traction forces 1 hour after Blebbistatin washout. (G) Total force as a function of time after removal of blebbistatin for GFP-palladin (black) and Palld4 (grey) cells. Each data point is an average of forces from N = 10 cells from independent experiments for each condition. The first data point represents the initial (pre-blebbistatin) force. The graphs show the increase in cellular traction forces as the cell recovers from blebbistatin washout, subsequent to 30 min incubation in Blebbistatin. (H) The percentage force (with respect to original forces) during recovery from blebbistatin after washout plotted as a function of time for EGFP-palladin cells (black) and Palld4 cells (grey). (I) The percentage increase of stress after washout of blebbistatin quantified as the difference between force recovered 1 hour after washout, Frecov and the force after incubation in blebbistatin for 30 minutes, Fblebb (with respect to the initial force prior to blebbistatin addition). The data represents an average for 20–30 cells of each type. (p < 0.05, Wilcoxon ranksum test).
