cDNA for gene expression quantifications was synthesized using a high-capacity cDNA Reverse Transcription Kit (Applied Biosystems, Milan, Italy), following the manufacturer’s instructions.
qRT-PCR was performed in an ABI-PRISM 7900HT Sequence Detector (Applied Biosystems, Milan, Italy) using the relative quantification method (2-ΔΔCt method), as previously described [10 (link)]. The genes were analyzed using the following TaqMan assays: SLC7A5 (Hs00185826_m1); SLC7A8 (Hs00794796_m1); PNMT (Hs00160228_m1); YAP1 (Hs00371735_m1); TNKS1 (Hs00186671_m1); CTNNB1 (Hs00355045_m1); AXIN2 (Hs00610344_m1); MCT8 (Hs00185140_m1); and FZD8 (Hs00259040_s1), all from Applied Biosystems.
Levels of mRNA expression were calculated with the Sequence Detection Software rel. 2.4 (Applied Biosystems, Milan, Italy) using the 2-ΔΔCt method and normalized to the housekeeping gene ACTB (Hs99999903_m1). All real-time reactions, including no template controls, were run in triplicate.
qRT-PCR was performed in an ABI-PRISM 7900HT Sequence Detector (Applied Biosystems, Milan, Italy) using the relative quantification method (2-ΔΔCt method), as previously described [10 (link)]. The genes were analyzed using the following TaqMan assays: SLC7A5 (Hs00185826_m1); SLC7A8 (Hs00794796_m1); PNMT (Hs00160228_m1); YAP1 (Hs00371735_m1); TNKS1 (Hs00186671_m1); CTNNB1 (Hs00355045_m1); AXIN2 (Hs00610344_m1); MCT8 (Hs00185140_m1); and FZD8 (Hs00259040_s1), all from Applied Biosystems.
Levels of mRNA expression were calculated with the Sequence Detection Software rel. 2.4 (Applied Biosystems, Milan, Italy) using the 2-ΔΔCt method and normalized to the housekeeping gene ACTB (Hs99999903_m1). All real-time reactions, including no template controls, were run in triplicate.
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