Quantitect hs actb assay

Total RNA was isolated with RNEasy Mini Kit (Qiagen, Hilden, Germany). The concentration and quality of RNA were evaluated using the RNA 6000 Nano LabChip Kit (Agilent 2100 Bioanalyzer, Agilent Technologies Inc., Santa Clara, CA, USA). Reverse transcription of total RNA was carried out using the IScript cDNA Synthesis Kit (Bio-Rad, Hercules, CA, USA), according to the manufacturer’s recommendations. Reverse transcription was performed using the IScript cDNA Synthesis Kit (Bio-Rad, Hercules, CA, USA). The relative mRNA expression levels of Nrf2, HO-1, and GCLC subunit were performed in triplicate using the QuantiTect Primer Assay and QuantiTect SYBR Green PCR Kit (Qiagen) on the MyiQ Thermal Cycler (Bio-Rad), as previously described [38 (link)]. QuantiTect Hs-ACTB Assay (Qiagen) was used as the normalizer. Normalized gene expression levels were given as the ratio between the mean value for the target gene and that of the β-actin in each sample.

Total RNA was isolated with RNEasy Mini Kit (Qiagen, Hilden, Germany). The concentration and quality of RNA were evaluated using the RNA 6000 Nano LabChip Kit (Agilent 2100 Bioer, Agilent Technologies Inc., Santa Clara, CA, USA). Reverse transcription of total RNA was carried out using IScript cDNA Synthesis Kit (Bio-Rad, Hercules, CA, USA) according to the manufacturer’s recommendations. The relative mRNA expression levels of Nrf2, HO-1 and GCLC were performed in triplicate using the QuantiTect Primer Assay and QuantiTect SYBR Green PCR Kit (Qiagen) on the MyiQ Thermal Cycler (Bio-Rad). QuantiTect Hs-ACTB Assay (Qiagen) was used as normalizer. Normalized gene expression levels are given as the ratio between the mean value for the target gene and that for the beta-actin in each sample.