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One g of RNA was reverse transcribed into cDNA using the Transcriptor First Strand cDNA synthesis kit (Roche, Indianapolis, IN, USA) and subjected to quantitative PCR using the SYBR-Green Supermix (Roche, Indianapolis, IN, USA) with the ABI Stepone Plus real-time PCR detection system (Applied Biosystems, Foster City, CA, USA)

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In another study, free fatty acids (FFA) were shown to cause circadian misalignment in hepatic HepG2 cells [43]