2.10 Quantitative real-time PCR analysis
Total RNA was extracted from cells using TRIzol reagent (Invitrogen), according to the manufacturer’s instructions. RNA was reverse transcribed into cDNA using a reverse transcription system (Vazyme, Nanjing, China). Real-time PCR was performed using a LightCycler® 480 System (Roche) with AceQ Universal SYBR qPCR Master Mix (Vazyme Biotechnology, Nanjing, China). The 2−ΔΔCT method was used to calculate the relative mRNA levels. GAPDH was used as an internal reference. The primer sequences used for quantitative real-time PCR were: TGF-β forward 5′-CCACCTGCAAGACCATCGAC-3′ and reverse 5′-CTGGCGAGCCTTAGTTTGGAC-3′.
Arg-1 forward 5′-CTCCAAGCCAAAGTCCTTAGAG-3′ and reverse 5′-AGGAGCTGTCATTAGGGACATC-3′, GAPDH forward 5′-AACGACCCCTTCATTGAC-3′ and reverse 5′-TCCACGACATACTCAGCAC-3′.