2.10 RNA isolation, reverse transcription and RT-PCR
Total RNA was extracted from skin lesion using a RNAprep Tissue Kit (TianGEN, China) and resolved in RNase-free water. The purity of total RNA was detected by nanodrop 2000 spectrophotometer(Thermo Fisher Scientific, USA). After reverse-transcript into complementary RNA with EvoScript Universal cDNA Master (Roche, USA), Real-time PCR was then performed by using FastStart Universal SYBR Green Master (ROX) (Roche, USA) on ViiA 7 Dx (Applied Biosystems, USA). Primers were shown as: TGFβ1, Forward: 5’-CCA TCG ACA TGG AGC TGG TGA AAG G-3’, Reverse: 5’-GCA GTG AGC ACT GAA GCG AAA GCC C-3’; IL-23 p19, Forward: 5’-TGG GCA TCT GTT GGG TCT-3’, Reverse: 5’-TGG GCA TCT GTT GGG TCT-3’. GAPDH as the control. All experiments were conducted according to the manufacturer’s instructions.