Quantitative Analysis of GGT1 Expression in Thymomas

Frozen specimens of thymomas and normal tissues were slightly shredded with scissors and total mRNA was extracted using TRIzol RNA Isolation Reagent (Gibco, #10296028). Expression level of GGT1 was analyzed by one-step probe qPCR using Luna Universal Probe One-Step RT-qPCR Kit (NEB, #E3006) and IDT PrimeTime^® qPCR Assays composed of Forward/Reverse primer pair and PrimeTime Probe. Primers were hGGT1 for GGT1 expression, and hGAPDH and hHPRT1 as internal standards. Primer and probe information is described in Supplementary Table S2. The mixture information is shown in Supplementary Table S3. The samples were centrifuged and each sample was measured by triplicate using a LightCycler^®480 System II (F. Hoffmann-La Roche Ltd, Basel, Switzerland). The mRNA expression levels of GGT1 were expressed relative to internal standards and compared between tumor and normal.