Figure 1
Involvement of NADPH oxidase in TP-induced ROS generation. (a) A representative sample for three independent FACS analyses is displayed (Left). Cells were treated with 500 μM thymidine for 48 h, and then ROS levels were measured in the cells. Proportions of cells in the M1 fraction are shown as relative levels of ROS. Relative ROS levels in KB/CV and KB/TP cells in the absence or presence of 500 μM thymidine are measured by using H2DCF-DA fluorescence dye (Right). (b) ROS levels in KB/CV and KB/TP cells incubated in the absence or presence of thymidine at 250 or 500 μM for 48 h. (c) Attenuated expression of IL-8 mRNA in KB/TP cells treated with NAC (left) and EUK8 (right) for 48 h. Expression of IL-8 in KB/CV and KB/TP cells was determined by real-time PCR. (d) Decreased expression of HO-1 and IL-8 mRNAs in KB/TP cells treated with apocynin for 48 h. (e) Expression of NOX isoforms in KB cells. Expression levels of NOX isoforms in KB/CV and KB/TP cells were determined by real-time PCR. (f) The effect of down-regulation of NOX2 and p22phox on the production of ROS in KB/CV and KB/TP cells. The cells transfected with NOX2 siRNA or p22phox siRNA were treated with 10 μM H2DCF-DA for 1 h and ROS levels were determined by using FACScan. (g) KB cells transfected with NOX2 siRNA or p22phox siRNA were treated with or without 500 μM thymidine for 48 h and IL-8 mRNA expression levels were determined by real-time PCR. Data are presented as mean ± SD. ∗P < 0.01.
