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Total RNA was harvested using TRIzol (Thermo Fisher), DNAse-treated (RQ1; Promega), and reverse-transcribed with the SuperScript III reverse-transcriptase (Invitrogen) using random hexamers (Thermo Fisher) as per manufacturer’s instructions. For quantitative RT-PCR, cDNAs were analyzed using an ABI 7500 Real-time PCR system (Applied Biosystems) using the Luna Universal qPCR Master Mix (NEB). Results were normalized to HPRT. The 2–ΔΔCt method was used to derive change in gene expression.
Copyright and License information: ©2022 Lemay et al
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