Figure - PMC

Skip to main content

View full-text article in PMC

. Author manuscript; available in PMC: 2011 May 12.

Published in final edited form as: Brain Res. 2010 Mar 18;1330:131–141. doi: 10.1016/j.brainres.2010.03.034

Rapid stretch injury-induced levels of reactive oxygen and nitrogen species in PC12 cells. (A) ROS and (B) NO were measured fluorimetrically at 0, 0.5, 1, 3, 6, 12 and 24 hours after RSI. Cells (1 × 10⁶/ml) were incubated with 5 μM CM-H₂DCFDA or 3 μM DAF-FM in 1 ml DPBS without Ca²⁺ and Mg²⁺ at 37 °C for 30 min for ROS or NO measurement, respectively. The level of ROS was calibrated by F_max, which is the fluorescence with the presence of 300 μM tert-butyl hydroperoxide, and was normalized to the fluorescence of PC12 cells prior to serum starvation as the control. The level of NO was normalized to total cell numbers. Data are represented as mean ± SD (n = 3, p < 0.0005 for ROS, and p < 0.01 for NO by one-way ANOVA).

Rapid stretch injury-induced levels of reactive oxygen and nitrogen species in PC12 cells. (A) ROS and (B) NO were measured fluorimetrically at 0, 0.5, 1, 3, 6, 12 and 24 hours after RSI. Cells (1 × 10⁶/ml) were incubated with 5 μM CM-H₂DCFDA or 3 μM DAF-FM in 1 ml DPBS without Ca²⁺ and Mg²⁺ at 37 °C for 30 min for ROS or NO measurement, respectively. The level of ROS was calibrated by F_max, which is the fluorescence with the presence of 300 μM tert-butyl hydroperoxide, and was normalized to the fluorescence of PC12 cells prior to serum starvation as the control. The level of NO was normalized to total cell numbers. Data are represented as mean ± SD (n = 3, p < 0.0005 for ROS, and p < 0.01 for NO by one-way ANOVA).