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.