RNA Sequencing of wound tissue of dbdb mice treated with PBS
Wound tissues were collected from db/db mice on POD6 in the PBS and MFGE8-pretreated fibroblast groups. Total RNA was extracted from these wound tissues using TRIzol Reagent, with three biological replicates per group (n = 3/group). RNA concentration and quality were assessed with a NanoDrop spectrophotometer. Three micrograms of total RNA were used for library preparation. Poly(A)+ mRNA was enriched with poly-T oligoattached magnetic beads, fragmented, and reverse-transcribed to generate first- and second-strand cDNA. After end repair, A-tailing, and adaptor ligation, cDNA fragments of 400500 bp were selected using the AMPure XP system (Beckman Coulter, USA). Libraries were amplified by 15 cycles of PCR, purified, and quantified using an Agilent Bioanalyzer 2100. Sequencing was performed on the NovaSeq 6000 platform (Illumina) by Shanghai Personal Biotechnology Co., Ltd.