Research Paper Volume 11, Issue 17 pp 7242—7256

Reactive oxygen species-mediated senescence is accelerated by inhibiting Cdk2 in Idh2-deficient conditions

Figure 5. Idh2 deficiency-mediated reactive oxygen species (ROS) generation activates p21 expression. (A) Western blot analysis of pro-inflammatory mediators and oxidative stress marker proteins in control and Idh2-knockdown mouse embryonic fibroblasts (MEFs). The following antibodies were used for detection: anti-Idh2, anti-iNOS, anti-Cox-2, anti-Prx-SO3, and anti-β-actin. (B) Western blot analysis of pro-inflammatory mediators and oxidative stress marker proteins between wild type and Idh2 knockout MEFs. The following antibodies were used for detection: anti-Idh2, anti-iNOS, anti-Cox-2, anti-Prx-SO3, and anti-β-actin. (C) Control and Idh2-knockdown MEFs were incubated with DCF-DA for 15 min at 37°C and intracellular ROS levels were analyzed by flow cytometry. (D) Wild type and Idh2 knockout MEFs were incubated with DCF-DA for 15 min at 37°C and intracellular ROS levels were analyzed by flow cytometry. (E) After transfecting MEFs with siIdh2, H2O2 was added for 3 days. NAC was added 4 h before H2O2 treatment. Western blot analysis was detected with the following antibodies. Data are expressed as means ± SD (n = 3). *p < 0.05, **p < 0.01, and ***p < 0.001.