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RT was performed by using MMLV (Moloney murine leukemia virus) RT kit (Invitrogen, Carlsbad, CA, USA) and reverse primer 827R (nt 1777-1800) (Table) according to manufacturer's recommendations. Total RNA was reverse transcribed with MMLV reverse transcriptase and oligodT primers (PerkinElmer Inc. Alternatively, a two-step RT-PCR amplification with the Vgen primer (5' AGCAAAAGCAGG) with MMLV reverse transcriptase (Promega, Madison, WI) and PCR with gene-specific primers was performed. |
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