Figure 4
Knock-down of Nox-1 or Nox-4 confirmed the involvement of NADPH oxidase in ROS generation after wound. (A) Depletion of either Nox-1 or Nox-4 significantly inhibited ROS production. After indicated plasmids were electroporated cells were scratched and growth factors were added as indicated. Thirty min later, cells were harvested and subjected to FACS analysis after DCF-DA staining. Representative data from two independent experiments are shown (right panel), and the extent of knock-down of Nox-1 and Nox-4 was measured by western blotting and RT-PCR, respectively (left lower panel). Representative FACS profiles are overlaid to reveal the differences between samples. Only the differences between samples with both wound and growth factor treatment were shown (left upper panel). Con shRNA: pSuper plasmid itselfplasmid transfected cells. (B) HaCaT cell migration was inhibited by depletion of either Nox-1 or Nox-4. Cells harboring indicated shRNAs were subjected to scratch wound assay. Data are presented as mean ± SD of two independent experiments (**; P < 0.01 compared to pSuper-transfected cells by student's t-test).
