Real-Time Quantitative PCR
To extract total RNA from monocytes, a RNeasy Mini Kit (Qiagen) was used. Once obtained RNA, cDNA was synthesized starting off from 0.5 μg of total RNA using high-capacity cDNA reverse transcription kit (Applied Biosystems). For real-time qPCR (real-time quantitative polymerase chain reaction), a SYBR Green master kit (Applied Biosystems) was used to evaluate PPAR-γ and β-actin mRNA relative levels in monocytes. Relative PPAR-γ mRNA expression was normalized against endogenous gen β-actin mRNA. The amounts of PPAR-γ mRNA relative to control were calculated with ΔCT method.