Reverse Transcription and Real-time PCR Analysis
Total RNA was extracted from snap-frozen skin tissues or HaCaT cells using the Trizol Reagent (Thermo Fisher Scientific, 15596026) on ice in an RNAse free environment, after which complementary DNAs were synthesized using the HiScript III 1st Strand cDNA Synthesis Kit (+gDNA wiper) (Vazyme Biotech, R312-01). The qRT-PCR was performed using the Step One Plus Real-Time PCR system (Applied Biosystems). β-Actin was used as the reference gene for normalization. The fold changes of mRNA relative expression were calculated using the comparative cycle method (2−ΔΔCt). The primer sequences are shown in Supplementary Table S1.