G3_1
After 3 days of implanting hydrogels of G3 group (-30% PSF and -40% KSF) , hydrogels were harvested from the defect and total RNA was extracted. mRNA enrichment was performed on total RNA using oligo(dT)-attached magnetic beads. The enriched mRNA with poly(A) tails was fragmented using a fragmentation buffer, followed by reverse transcription using random N6 primers to synthesize cDNA double strands. The quality and purity of RNA were further ascertained by ShenZhen BGI Genomics Co., Ltd.. The constructed libraries were quality-checked and sequenced after passing the quality control (https://biosys.bgi.com). Sequencing was performed on DNBSEQ platform with PE150 (read length). Gene expression quantification was performed using RSEM (v1.3.1) software, and gene expression clustering heatmaps across different samples were generated using pheatmap (v1.0.8) . DESeq2 (v1.4.5) (or DEGseq or PoissonDis ) was employed for differential gene detection, with criteria set as Q value0.05 or FDR0.001.