Figure 6.

Overexpression of Magmas is associated with elevated enzymatic activity of antioxidants and mitochondrial DNA copy number. (a) For the quantitative measurement of mtDNA, isolated mitochondria expressing a basal level of Magmas (UT) and higher amount of protein (Mag ↑) were stained with SYBr green and the fluorescence emitted upon its binding to mtDNA was represented as fold fluorescent at 520 nm (F₅₂₀) over the control (UT). (b) To quantify the mtDNA copy number, two regions of mtDNA, namely ND1 and CO2, were amplified by qPCR and data are presented as fold change in the copy number upon Magmas overexpression over untransfected control. (c and d) Level of cellular GSH (c) and GSSG (d) was measured in cells overexpressing Magmas in a luminescence-based assay in the presence or absence of oxidative stress. (e) The enzyme activity of glutathione peroxidase was measured spectrophotometrically under untreated condition and after treatment of 25 μM NaAsO₂. (f) Isolated mitochondria were utilized to measure the activity of MnSOD upon Magmas overexpression. The activity is presented in terms of rate of the reaction. Data represented as mean±S.E.M., n=3; *P<0.01 (two tailed), **P<0.001 (two tailed) and ***P<0.0001 (two tailed)