Real-time PCR
For each sample, total RNA was isolated from 50 mg of frozen LM
samples using the RNeasy Fibrous Tissue kit (QIAGEN GmbH, Hilden,
Germany), according to the manufacturer's instructions. Total RNA
was quantified by absorbance at 260 nm, and the integrity of total
RNAwas checked by agarose gel electrophoresis and ethidiumbromide
staining of the 28S and 18S
bands.