Journal: Cancers

Article Title: CPT1A Over-Expression Increases Reactive Oxygen Species in the Mitochondria and Promotes Antioxidant Defenses in Prostate Cancer

doi: 10.3390/cancers12113431

Figure Lengend Snippet: Mitochondrial reactive oxygen species (ROS) are increased in CPT1A-OE cells. ( A ) Electron paramagnetic resonance spectroscopy (EPR) traces of the four cell lines tested, including the mitochondrial probe by itself. Amplitude and linewidth provide information of the amount of ROS. The concentration was acquired by Spin Fit followed by SpinCount module (Bruker). ( B ) Quantification of the EPR traces normalized to protein content. *** p < 0.001. ( C ) Gene expression analysis of OE versus KD cells for SOD1 ns), SOD2 ( p = 6.9 × 10 −4 ), and SOD3 ( p = 0.015). Only SOD2 is a mitochondrial enzyme. ( D , E ) Western blots of SOD2 expression after incubation with fatty acids (Oleic and palmitate mixture; 25 µM each) in FBS ( D ) or charcoal stripped serum (CSS) ( E ) for 48 h. ( F ) Schematic of the role of SOD2 in metabolizing superoxide and the role of glutathione in eliminating H 2 O 2 . GPX = Glutathione peroxide. GSSG = oxidized glutathione.

Article Snippet: Antibodies: CPT1A: 15184-1-AP, (Proteintech, Rosemont, IL, USA); GAPDH: CST 5174, (Cell Signaling Technology, Beverly, MA, USA); SOD2: CST 13141, Cell Signaling Technology.

Techniques: Electron Paramagnetic Resonance, Spectroscopy, Concentration Assay, Gene Expression, Western Blot, Expressing, Incubation