Quantitative real-time polymerase chain reaction (qRT-PCR) analysis of enzymic antioxidant (SOD1, SOD2, SOD3, CAT1, GPX1, GPX2, GPX3, GPX4, GPX5, GSR, GSS, GSTZ1) gene expression was performed as previously described [40]. DPSCs at selected PDs throughout their proliferative lifespans were cultured, RNA extracted, and cDNA synthesized, as above. cDNA amplification was performed using the Applied Biosystems™ ViiA™ 7 Real-Time PCR System and TaqMan® primers (ThermoFisher Scientific, Supplementary Table S2), according to the manufacturer’s protocols. qRT-PCR was performed MicroAmp™ Fast Optical 96-Well Reaction Plates (ThermoFisher Scientific), per the manufacturer’s instructions. Relative fold changes in enzymic antioxidant gene expression (RQ) were calculated using the 2–ΔΔCt method [41], normalized versus an 18S rRNA housekeeping gene.

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.