*4.4. Quantitative PCR for Relative Telomere Length Assessment*

Average relative telomere length was measured as previously described [2,28,31,32] with modifications. IFNB1 was used as a single copy gene as published by Vasilishina et al (primer sequences detailed in Table A7). qPCR reactions mixes were prepared according to Table A8 with a standard curve performed for each run. qPCR run in LightCycler 480 II (Roche Diagnostics International Ltd, Rotkreuz, Switzerland) under following conditions: pre-incubation at 95 ◦C for 10 min, 35 cycles of 95 ◦C for 15 s, 60 ◦C for 60 s, and 72 ◦C for 10 s, followed by melting curve at 95 ◦C for 5 s, and 65 ◦C for 60 s. Triplicates for each sample were performed and concentration of sample calculated according to same-run standard curve and averaged for each sample. Relative telomere length (T/S ratio) of each sample was calculated as the ratio between average concentration of telomere reactions to the average concentration of the single copy gene reactions.
