Fig. 6: CDS2 interacts with OPA1 at the VAL-280 site.

A Co-IP Coomassie Bright Blue Dyeing Experiment. B Mass spectrometry analysis to obtain mitochondrial-related protein OPA1 with a high comprehensive score. C HUVECs were subject to immunoprecipitation (IP) with an anti-CDS2 antibody. The immunoprecipitation complex was probed via western blotting using an anti-OPA1 antibody (n = 3 per group). D HUVECs were treated with si-METTL3 and were then subjected to immunoprecipitation (IP) with an anti-CDS2 antibody. Co-IP of OPA1 was detected by western blotting using an anti-OPA1 antibody (n = 3 per group). E Protein-Protein Interaction Networks (PPI) predicted the binding sites of CDS2 and OPA1. F–H HUVECs transfected with CDS2-HA or CDS2 mutants for 24 h, followed by immunoprecipitation. Detection of CDS2 (wild type and mutants) was performed via Western blotting using an anti-HA antibody (n = 3 per group). I, J, and K Assessment of tubular structures and ROS in HUVECs under various conditions to evaluate cellular function and metabolism (n = 3 per group). L ATP assay conducted to assess the energy metabolism of HUVECs in different environmental conditions (n = 3 per group). M Western blot was utilized to detect the levels of proteins OPA1, METTL3, CDS2, and COX IV in HUVECs across various environments in each experimental group (n = 3 per group). N, O, and P Observation and analysis of the healing process and the expression of KRT14 in mouse back skin across different environments, accompanied by blood flow imaging to assess vascular changes (n = 4 per group). The data are the means ± SD of at least three independent experiments, *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 are considered significant.