qPCR Analysis of Skin Marker Genes

RNA extraction and cDNA synthesis were conducted as previously described [16 (link), 26 (link)]. Real-time reverse transcriptase polymerase chain reaction (RT-PCR) was performed with the LightCycler 480 Instrument II Real-Time PCR System (Roche Diagnostics Ltd., Rotkreuz, Switzerland). The relative expression of each mRNA was normalized to β-actin as an internal control. The primers used in this study were as follows: IVL: forward 5′-TCCTCCAGTCAATACCCATCAG-3′; reverse 5′-CAGCAGTCATGTGCTTTTCCT-3′; CK13: forward 5′-CCCCAGGCATTGACCTGAC-3′; reverse 5′-GTGTTGGTAGACACCTCCTTG-3′; ACTB: forward 5′-TTGTTACAGGAAGTCCCTTGCC-3′; reverse: 5′-ATGCTATCACCTCCCCTGTGTG-3′.