KLF2/PPAR? axis contributes to trauma-induced heterotopic ossification by regulating mitochondrial dysfunction

Trauma-induced heterotopic ossification (HO) is one of the most complex disorders after musculoskeletal injury and is characterized by aberrant extraskeletal bone formation. Recent studies have shed light on the critical role of dysregulated osteogenic differentiation in aberrant bone formation. Krupel-like factor 2 (KLF2) and peroxisome proliferator-activated receptor gamma (PPAR?) are master adapter proteins that link cellular responses to osteogenesis; however, their roles and relationships in HO remain elusive. Overall design: Whole transcript genome sequencing was performed on the soft tissue at the Achilles tendon (provided by Shanghai OE Biotech. Co., Ltd). The steps are as follows: total RNA was extracted and ribosomal RNA was digested using ribo-Zero kit. After interrupt reagent was added to break the RNA into short fragments, and a chain was synthesized with random primer of six bases using the interrupted RNA as template cDNA, and then prepared a two-strand synthesis reaction system to synthesize two-strand cDNA, dUTP instead of dTTP in the synthesis of cDNA, and then one strand containing dUTP was digested by UNG enzyme method, and only the cDNAs of the different junctions of the strand were retained. A chain used kit to purify a strand of cDNA, a strand of purified cDNA was then repaired at the end, an a-tail was added, and sequenced.