RNA-seq of ATDC5 chondrocytes treated with asperuloside under TNF-α and IL-1β stimulation compared with PBS-treated and untreated controls
RNA-seq of ATDC5 chondrocytes treated with asperuloside under TNF-α and IL-1β stimulation compared with PBS-treated and untreated controls
ATDC5 cells were cultured under standard conditions (37°C, 5% CO₂) in DMEM/F12 medium supplemented with 5% FBS and antibiotics.  Cells were divided into three groups: untreated control, TNF-α/IL-1β–stimulated group (OA model), and TNF-α/IL-1β–stimulated group treated with asperuloside (ASP).  After 24 hours of treatment, cells were harvested for RNA extraction.  Total RNA was extracted from ATDC5 cells using TRIzol reagent (Invitrogen) according to the manufacturer's instructions.  RNA concentration and purity were assessed using a NanoDrop spectrophotometer, and RNA integrity was confirmed with an Agilent 2100 Bioanalyzer (RIN ≥ 8.0).  RNA-seq libraries were prepared from 1 µg of total RNA using the Illumina TruSeq Stranded mRNA Library Prep Kit (Illumina, USA) following the manufacturer's protocol.  After adapter ligation and PCR amplification, libraries were quantified and validated using a Qubit fluorometer and Agilent Bioanalyzer. 