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Quantitative analysis of mRNA expression was performed using the PerfeCTa SYBR Green SuperMix Kit (Quantabio, Beverly, MA, USA) on a CFX96 Dx Real-Time PCR Detection System (Bio-Rad, Hercules, CA, USA) following manufacturer’s instructions. Expression of mRNA was evaluated through standard curves generated for each target gene by plotting Ct values versus log cDNA dilution. UBC was used to normalize mRNA levels. PCR products were verified by a melting profile and agarose gel electrophoresis to rule out nonspecific PCR products and primer dimers.
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Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
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