Panaxatriol exerts anti-senescence effects and alleviates osteoarthritis and cartilage repair fibrosis by targeting UFL1

Osteoarthritis is the most commmon degenerative joint disease which can lead to disability eventually. However, there are currently no safe and effective interventions available. Therefore, it is urgent to develop effective drugs that can reduce cartilage damage and treat OA.With the in vitro culturing of human cartilage explants and mouse OA model, it was found that, panaxtriol, a natural small molecule drug, could promote chondrocyte anabolism and inhibit catabolism. It could also reduce the loss of cartilage matrix , decrease the subchondral osteosclerosis, and relieve pain in mice. Eventually, it could delay the progression of OA. Subsequently, the binding target of panaxtriol was found to be UFL1 through the target stability of drug affinity reaction (DARTS). The UFL1 knockout cell line was constructed using CRISPR-Cas9. Then, the regulation of chondrocyte metabolism by panaxtriol was found to dependent on UFL1. Transcriptome sequencing of UFL1 knockout cells were conducted. Throug..., RNA sequencing: C28/I2 cells and UFL1 knock-out cells were send to RNA sequencing analysis, each group has 3 repeats. According to the technology vendor's process, RNA integrity was assessed using the RNA Nano 6000 Assay Kit of the Bioanalyzer 2100 system (Agilent Technologies, CA, USA). After the construction of the library, the library was initially quantified by Qubit2.0 Fluorometer, then diluted to 1.5ng/ul, and the insert size of the library is detected by Agilent 2100 bioanalyzer. After the library is qualified, the different libraries are pooling according to the effective concentration and the target amount of data off the machine, then being sequenced by the Illumina NovaSeq 6000. Raw data (raw reads) of fastq format were firstly processed through fastp software to get clean data with high quality. We selected Hisat2 as the mapping tool to read mapping to the reference genome. FeatureCounts (v1.5.0-p3) was used to count the reads numbers mapped to each gene. And then FPKM of ea..., , **Panaxatriol exerts anti-senescence effects and alleviates osteoarthritis and cartilage repair fibrosis by targeting UFL1**

[https://doi.org/10.5061/dryad.8cz8w9gzp](https://doi.org/10.5061/dryad.8cz8w9gzp)

**Quantification of gene expression level** feature Counts v1.5.0-p3 was used to count the reads numbers mapped to each gene. And then FPKM of each gene was calculated based on the length of the gene and reads count mapped to this gene. FPKM, expected number of Fragments Per Kilobase of transcript sequence per Millions base pairs sequenced, considers the effect of sequencing depth and gene length for the reads count at the same time, and is currently the most commonly used method for estimating gene expression levels.

**Differential expression analysis** Differential expression analysis of two conditions/groups (three biological replicates per condition) was performed using the DESeq2 R package (1.20.0). DESeq2 provide statistical routines for determining differential expression ...,