Power sybr green 1 dye chemistry

1 µg RNA samples was reverse transcribed with the SuperScript™ First-Strand Synthesis system and random hexamers as primers (Life Technologies, Carlsbad, CA, USA). The expression levels of FXN, NRF2, NQO1, HO-1, and GCL were measured by qRT-PCR in an ABI PRISM 7500 Sequence Detection System (Life Technologies) using Power SYBR Green I dye chemistry (ThermoFisher Scientific, Walthman, MA, USA). Data were analyzed using the 2^−∆∆Ct method with TBP (TATA box binding protein) and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as housekeeping genes, and data are shown as fold change relative to controls. Primers used for qRT-PCR are reported in Petrillo et al. [74 (link)].

Total RNA was extracted from leukocytes using Total RNA Purification Plus Kit (Norgen, Biotek Corp., Thorold, ON, Canada), according to manufacturer’s protocol. RNA quantification was performed on a NanoDrop2000 Spectrophotometer (Thermo Scientific, Waltham, MA, USA). The purity of RNA was assessed by measuring the ratio of absorbance at 260 nm and 280 nm. An amount of 1μg RNA was reverse transcribed with the SuperScriptTM First-Strand Synthesis system and random hexamers as primers (Life Technologies, Carlsbad, CA, USA). The expression levels of GCL, GPX4, and NRF2 were measured by qRT-PCR in an ABI PRISM 7500 Sequence Detection System (Life Technologies) using Power SYBR Green I dye chemistry (ThermoFisher Scientific, Walthman, MA, USA). Data were analyzed using the 2 ^∆∆ Ct method with TBP (TATA box binding protein) as housekeeping gene and expressed as fold change relative to controls. Primers used for qRT-PCR are reported in Table 2.