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Fig. 5. TERT overexpression decreases mitochondrial mass and mtDNA content, reverses expression of candidate genes for retrograde response and improves resistance against apoptosis. (A) Relative mitochondrial mass per cell (relative to that in MRC5 cells under normoxia) as measured by NAO fluorescence in flow cytometry. (B) Relative MtDNA copy number as measured by real-time PCR. All data are mean±s.e.m. from at least three experiments. Asterisks indicate significant differences between parental and TERT-overexpressing cells with P<0.05. (C-F) Relative mRNA expression levels of candidate marker genes for retrograde response (Passos et al., 2007) in senescent MRC-5 (SEN), young MRC-5 (YOU) and MRC5-TERT cells (TERT). Expression colour code is indicated at the bottom. (C) Ca²⁺-related signalling, (D) metabolism, (E) mitochondrial function and (F) stress response. (G) Caspase 3/7 activity (in arbitrary units) in MRC5 and MRC5-TERT cells treated for 2 hours with H₂O₂ at the indicated concentrations and assayed after 24 hours. Data are mean±s.e.m. from three experiments. (H) Caspase 3/7 activity (in arbitrary units) in MRC5 and MRC5-TERT cells treated for 48 hours with etoposide at the indicated concentrations and assayed after 3 days. Data are mean±s.d. from two experiments.

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