Quantitative Real-Time PCR (qRT-PCR) Analysis

Total RNA was isolated from MEFs using Trizol (Invitrogen) according to the manufacturer’s protocol. RNA quantity was determined by agarose gel electrophoresis and by spectrophotometry. Single-stranded complementary DNA (cDNA) was obtained from reverse transcription of 600 ng of RNA using RT-PCR kit (BD Biosciences). qRT-PCR was performed in triplicate using SYBR (Takara Bio, Japan). β-actin primers: 5ʹ-AGAGGGAAATCGTGCGTGAC −3ʹ, 5ʹ-CAATAGTGATGACCTGGCCGT −3ʹ; α-SMA primers: 5'-ATGCAGAAGGAGATCACAGC-3', 5'-CAGCTTCGTCGTATTCCTGT-3'; FAP primers: 5'-TTCTGCCTCCTCAGTTTGAC-3', 5'-CTGTGAGCTGGTCCTCAACT-3'; FSP1 primers: 5'-AGGGTGACAAGTTCAAGCTG-3', 5'-GCAGGACAGGAAGACACAGT-3'; Vimentin primers: 5'-TCCTCGAGCAGCAGAACAAAATCC-3', 5'-CAGGGCAGCAGTGAGGTC-3'; IL6 primers: 5'-TAGTCCTTCCTACCCCAATTTCC-3', 5'-TTGGTCCTTAGCCACTCCTTC-3'; CXCL12 primers: 5'-TGCATCAGTGACGGTAAACCA-3', 5'-CACAGTTTGGAGTGTTGAGGAT-3'; CXCL10 primers: 5'-CCAAGTGCTGCCGTCATTTTC-3', 5'-GGCTCGCAGGGATGATTTCAA-3'; CXCL1 primers: 5'-GCTTGAAGGTGTTGCCCTCA -3', 5'-CTATGACTTCGGTTTGGGTGC-3'; FGF10 primers: 5ʹ- CACATTGTGCCTCAGCCTTTC-3ʹ, 5ʹ- CCTCTATTCTCTCTTTCAGCTTAC-3ʹ; The relative expression level of each mRNA was analyzed by the 2^−ΔΔCt method. All experiments were repeated three times.