adck3 mutant cells are sensitive to H2O2 and display an increase in oxidative stress.

(A). Analysis of ADCK3 levels in control (C) and adck3 mutant (P1, P2 and P3) mutant fibroblasts. WCEs (30 μg) were separated via SDS-PAGE and immunoblotting performed for ADCK3 and PCNA as a loading control. (B). Analysis of CoQ levels in adck3 mutant cells. Total cellular levels of CoQ10 and CoQ9 were determined by RP-HPLC, normalised to non-esterified cholesterol and mitochondrial mass as described in Materials and Methods. (C, D). IR/H2O2 sensitivity of control and adck3 mutant cell lines. Control and adck3 mutant fibroblasts were exposed to various doses of H2O2 (C) or IR (D) and survival assessed via colony formation assays. AT1ABR is an Ataxia Telangiectasia (A-T) cell line used as a positive control for IR sensitivity. n = 3. (E). Analysis of 4-HNE Michael adducts and nitro-Y levels in adck3 mutant cell lines. A representative example of the images obtained from this experiment are depicted. Fluorescence intensity values from individual cells were normalised to control values ± S. D. 50 cells analysed per experiment. n = 3. Statistical analysis was performed with Student’s t-test in all cases. * p p