Abi 9700 detection system

Trizol reagent (Tiangen, Beijing, China) was used for total RNA isolation of the skin tissue. cDNA was prepared via reverse transcription of RNA. cDNA, primers, and SYBR premix (Roche, Basel, Switzerland) were combined on qRT-PCR using the ABI 9700 Detection System (Thermo Fisher Scientific, Waltham, MA), and GAPDH was used as control. Supplemental Table S1 shows the primers. All experiments were repeated three times. qRT-PCR for miRNA was done using miScript SYBR Green PCR kit (Qiagen, Hiden, Germany) according to the manufacturer’s instruction. All results were normalised to U6 small RNA levels measured using the Hs_RNU6B_2 miScript Primer Assay kit (Qiagen). Supplemental Table S1 shows the primers. All experiments were repeated three times.

Total RNA was isolated from tissues, cells, or hAFSC-exo using TRIzol Reagent (Sigma, United States) and reverse-transcribed into cDNA using the PrimeScript RT reagent kit (Takara, Japan). qRT-PCR was performed on an ABI 9700 Detection System (Thermo Fisher Scientific, United States). The relative expression levels of the target genes were calculated using the 2^–ΔΔCt method. GAPDH and U6 were used as controls for the mRNA and miRNA reactions, respectively. Supplementary Table 2 lists the primers. All experiments were repeated three times.

Total RNA was isolated from healing skin, HDFs, and UCB-MSC-exo using TRIzol reagent (Sangon Biotech Co., Ltd., Shanghai, China). cDNA was synthesized from total RNA using the cDNA Synthesis Kit (Takara, Japan). qRT-PCR analysis was performed using SYBR Green Master (Roche, Switzerland) in an ABI 9700 Detection System (Thermo Fisher Scientific, MA, USA). GAPDH mRNA was used as an internal control. qRT-PCR for miRNA was performed using the Bulge-Loop^TM miRNA qRT-PCR Starter Kit (Ribobio, China) according to the manufacturer’s instructions. U6 small RNA was used as an internal control. Table S2 lists the primers used. All experiments were repeated in triplicate.