Research Paper Volume 11, Issue 17 pp 7242—7256

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

Figure 7. Overexpression of Idh2 prevents the acceleration of senescence in mouse embryonic fibroblasts (MEFs). (A) SA-β-gal staining of control and Idh2-overexpressing MEFs. pLenti 6.3 Idh2 plasmid was transfected into Passage 8 (P8) MEFs. (B) Statistical analysis of SA-β-gal-stained positive cells between control and Idh2-overexpressing MEFs. (C) Senescence-associated marker proteins were detected using western blot analysis in control and Idh2-overexpressing MEFs. The following antibodies were used for detection: anti-Idh2, anti-p16, anti-p21, anti-p53, and anti-β-actin. (D) Pro-inflammatory mediators, reactive oxygen species (ROS) marker proteins, and cyclin-dependent kinase 2 were detected using western blot analysis. The following antibodies were used for detection: anti-iNOS, anti-Cox-2, anti-Prx-SO3, and anti-β-actin. (E) Relative intracellular ROS levels were detected in control and Idh2-overexpressing MEFs. Intracellular ROS were detected by flow cytometry. Data are expressed as means ± SD (n = 3). *p < 0.05, **p < 0.01, and ***p < 0.001.