Platelet-derived mitochondria transfer accelerates fracture healing by promoting osteogenic differentiation and metabolic reprogramming of adipose-derived stem cells

Figure 5 H3K18 lactylation facilitates osteogenesis-related gene transcription. A: The heatmap showed the differentially expressed RNAs in adipose-derived stem cells (ASCs) in the control and mitochondria-treated groups; B: The volcano plot showed differentially expressed RNAs in ASCs in the control and mitochondria-treated groups; C: Gene Set Enrichment Analysis results showed the functions of differentially expressed genes; D: The bubble plot showed the Kyoto Encyclopedia of Genes and Genomes enrichment analysis of RNAs in different pathways; E-H: ASCs were treated with mitochondria and 2-deoxy-D-glucose. The expression of AXIN2, BMPR1B, COL1A1, and OSTN was measured using quantitative real-time polymerase chain reaction (E). Chromatin immunoprecipitation was conducted to determine H3K18 lactylation in the gene promoter (F). The transcriptional activity was evaluated by luciferase reporter assay (G). Protein levels of AXIN2, BMPR1B, COL1A1, and OSTN were examined by western blotting (H). n = 3. Data were analyzed by one-way ANOVA followed by Tukey’s post hoc test and are presented as mean ± SD. 2-DG: 2-deoxy-D-glucose.