Exosome Isolation from NLF and pHNEC

NLF was collected from NP and DNS patients, respectively, and detailed methods can be found in our previous work [11 (link)]. Exosomes were isolated by differential ultracentrifugation as we have described before [11 (link)]. Briefly, the supernatants from both NLF and pHNEC culture medium were centrifuged at 6,000 × g for 30 min and then 10,000 × g for 60min at 4°C, followed by ultrafiltration (0.2μm filter; Sarstedt, Nümbrecht-Rommelsdorf, Germany) and qEV size-exclusion columns (Izon Science, Christchurch, New Zealand). Thereafter, the supernatant was then ultracentrifuged at 100,000 × g for 60min at 4°C (Type 90 Ti Rotor; Beckman Coulter, Inc., Brea, CA, USA) to pellet the exosomes. The exosome pellets were then washed using PBS for cell experiments.