Fig. 2.

Cell damage and related cell death of cancer cells treated with elevated FSS. (A) Representative PI flow cytometry scatter plots to quantify cell membrane damage in DU145, PC3 and LNCaP cells. The percentage of the cell population in the undamaged cell gate (PI negative) is shown for each plot. (B) Percentage of undamaged cancer cells in populations treated with (shear stress) or without (static) 10 pulses of FSS. (C) Average number of PI-negative (undamaged) cancer cells treated with FSS, normalized to untreated static controls. (D) Normalized undamaged cancer cells as a function of FSS pulses, ranging from 0 to 10. (E) Uptake of FITC-tagged dextrans of different MW and hydrodynamic radii by cancer cells under static conditions as a measure of cell membrane damage. (F) Uptake of different dextrans by cancer cells treated with 10 pulses of FSS. Cell lines are color-coded as in E. (G) Representative flow cytometry scatter plots of 3000 MW FITC-tagged dextran/PI cell fate tracking. Cells positive for dextran indicate cell membrane damage, cells positive for PI represent dead cells, and cells negative for both indicate viable cells. The percentage of the cell population in each cell gate is indicated on the plots. (H) Proportion of PI-positive cells, normalized to to the corresponding undamaged dextran subpopulation, in dextran-negative and dextran-positive populations of cancer cells treated with 10 pulses of FSS. Data are presented as mean±s.d. N=3 independent experiments. *P<0.05, **P<0.01, ***P<0.005, ****P<0.001 (unpaired two-tailed t-test). ^###P<0.005 (simple linear regression to confirm significant deviation from zero).