Gene-specific PCRs were performed in a total volume of 25 μL containing 5 μL cDNA, 2.5 μL 10× PCR buffer (Mg2+ free), 0.5 μL 10 mM dNTP mix, 0.5 μL 10 μM primer (forward and reverse each), 0.75 μL 50 mM MgCl2, 0.1 μL (5 U/μL) Taq polymerase (all from Invitrogen), and H2O. The 25-μL PCR steps were 30 seconds of denaturation at 96°C, 30 seconds of annealing, and 45 seconds of extension at 72°C, followed by an end-extension step of 5 minutes at 72°C after the last cycle. The primers for Hsp47 were (forward, 5′-TTCTGCCTCCTGGAGGCG-3′; reverse, 5′-CGCTCAGCACTGCCTTGG-3′, position, 257-274; product size, 267; annealing temperature, 58.0°C), for Col1α1 were (forward, 5′-GATGGACTCAACGGTCTCC-3′; reverse 5′-CCTTGGGGTTCTTGCTGATG-3′; position, 3576-4034; product size, 458; annealing temperature, 57.0°C), and for GAPDH (forward, 5′-GAAGGTGAAGGTCGGAGTC-3′; reverse, 5′-GAAGATGGTGATGGGATTTC-3′; position, 6-231; product size, 225; annealing temperature, 57.0°C). The functionality of primers was tested on cDNAs obtained from different tissues before the experiments to exclude false-negative results (data not shown). The band intensity was measured in light units with an imaging workstation (Lumi-Imager; Roche, Mannheim, Germany). Quantification was performed with the accompanying software (Lumi-Analyst software, Roche). The final amount of PCR product was expressed as the ratio of the Hsp47 or the Col1α1 gene amplified to that of the GAPDH gene.