2.5. mRNA extraction and reverse transcription
Reagents were purchased from Thermofisher®. Trireagent was
obtained from Sigma® and ARN spin column were purchased from
Macherey Nagel® and used following manufacturer's recommendations.
The quantity and quality of RNA was assessed by absorbance
measurement at 260 and 280 nm. Then, 500 ng of each mRNA sample
were reverse transcribed using 20 U of RNasin (Promega®), 0.5 mM
of RNAse free dNTP and 200 U of MMLV (Moloney Murine Leukemia
Virus Reverse transcriptase Promega®).
2.6. Real-time PCR
The sequences of forward and reverse primers were designed using