Fig. 6. The modulation of Sox18 expression does not alter the proliferation of pulmonary artery endothelial cells.
PAEC transfected with either pCMV6-Sox18 or Sox18 siRNA for 24 h were trypsinized, seeded onto a 6-well plate at a density of 5.5 × 105 or 5.0 × 105 cells per well, respectively, and grown for an additional 48 h at 37°C in an incubator. After 48 h and 72 h post transfection, the cellular proliferation was evaluated by counting the cell number with a hemocytometer. The PAEC expressing pCMV6-Sox18 (A) or Sox18 siRNA (B) showed no difference in their cell count compared to their respective controls. Further, PAEC transfected with either pCMV6-Sox18 or Sox18 siRNA for 24 h were trypsinized, seeded onto a 6-well plate at a density of 5.5 × 105 or 5.0 × 105 cells per well, respectively, and grown for an additional 24 h at 37°C in an incubator. The cells were then exposed to 6 h or 12 h of shear (20 dyn/cm2), and the cellular proliferation was evaluated by counting the cell number with a hemocytometer. The PAEC expressing pCMV6-Sox18 (C) or Sox18 siRNA (D) showed no difference in their cell counts compared to their respective controls. To evaluate apoptotic cell death, PAEC were transfected for 48 h with either pCMV6, pCMV6-Sox18, scrambled siRNA, or Sox18 siRNA. The cells were then subjected or not to shear (20 dyn/cm2, 3 h). No differences in cellular apoptosis were observed with either Sox18 over-expression or Sox18 silencing in the presence or absence of shear (E). Values are mean ± SEM, n=3.