“…After 24 h, total RNA was extracted using TRIzol reagent (Invitrogen), and 1 μ g of RNA was converted into cDNA using the ImProm‐II Reverse Transcription System (Promega, Madison, WI, USA). Quantitative real‐time PCR procedures were performed using LightCycler 480 SYBR Green I Master (Roche Diagnostics, Mannheim, Germany), an equal amount of cDNA, and specific primers for genes encoding type I collagen and its modifying enzymes, as follows: collagen type I alpha‐2 chain ( COL1A2 ) , 5′‐TAATGGGGAAGCTGGATCTG‐3′ (forward) and 5′‐GTCCCTGAGCACCATTGTTT‐3′ (reverse); LH1 ( PLOD1 ) , 5′‐GAAGCTCTACCCCGGCTACT‐3′ (forward) and 5′‐CTTGTAGCGGACGACAAAGG‐3′ (reverse); LH2 ( PLOD2 ) , 5′‐GGGAGTTCATTGCACCAGTT‐3′ (forward) and 5′‐GAGGACGAAGAGAACGCTGT‐3′ (reverse); LH3: PLOD3 , 5′‐CACTACGGCCAGTGGTCAG‐3′ (forward) and 5′‐GTGGGCACATTCTCGTAGC‐3′ (reverse); GLT25D1 ( GLT25D1 ) , 5′‐GATGCTGCCTGTGGACGAGTTC‐3′ (forward) and 5′‐CTCACATAGCCATCGTCTCCTG‐3′ (reverse); LOX ( LOX ) , 5′‐CGCTGTGACATTCGCTACACAGGAC‐3′ (forward) and 5′‐CATTGGGAGTTTTGCTTTGCCTTCT‐3′ (reverse); and glyceraldehyde‐3‐phosphate dehydrogenase ( GAPDH ) , 5′‐GCTCTCCAGAACATCATCC‐3′ (forward) and 5′‐GTGTCGCTGTTGAAGTCAG‐3′ (reverse). Results were analyzed using the LightCycler 480 Instrument (Roche Diagnostics).…”