RNA-Seq of rat condylar chondrocytes: IL-1 inflammation & DFSCs-Exos intervention
Total RNA was extracted from chondrocytes of three experimental groups (MCC, Mcc+exo, mcc+IL-1b) using TRIzol reagent (Invitrogen) following the manufacturers protocol. RNA quality was verified by Nanodrop 2000 (Thermo Fisher) for purity (A260/A280 = 1.82.1), Agilent 2100 Bioanalyzer for integrity (RIN  8.0), and Qubit 4.0 for concentration. For mRNA library construction: Oligo(dT) beads were used to enrich poly(A)+ mRNA, which was fragmented into ~200bp fragments. First-strand cDNA was synthesized with random hexamers, followed by second-strand cDNA synthesis, end repair, A-tailing, and adapter ligation. Target fragments were amplified by PCR. For miRNA library construction: Small RNAs (~1830 nt) were isolated by polyacrylamide gel electrophoresis, ligated with 3 and 5 adapters, reverse-transcribed into cDNA, and amplified by PCR. Libraries were quantified by Qubit and Agilent 2100, then sequenced on an Illumina NovaSeq 6000 platform (paired-end 150bp for mRNA, single-end 50bp for miRNA).