Quantitative RT-PCR Analysis of CD44 Expression

Total RNA was isolated from the cells using the TRIzol reagent (Life Technologies). For the synthesis of cDNA, RT reactions were performed by incubating 200 ng of total RNA with a reaction mixture containing 0.5 μg/μL oligo dT_12–18 and 200 U/μL moloney murine leukemia virus RT (Life Technologies). Real-time RT-PCR was carried out using a Roche Light Cycler (Mannheim, Germany) with the Takara SYBR Premix ExTaq System for relative quantification. Primers were synthesized by Bioneer (Daejeon, Republic of Korea) and primer sequences for the human genes are described in our previous studies [56 (link), 57 (link)]. PCR primers for CD44 are 5′-AGCAGCACTTCAGGAGGTTAC-3′ and 5′-TGCCTCTTGGTTGCTGTCTC-3′.