Prompt Description

Figure creation: MCF7 cell culture. Seeding followed by treatment with five different cellular stressors: 1. Control, 2. Hypoxia (the flask lid was tightly sealed with parafilm), 3. UV Radiation, 4. Serum Depletion (0.5% FBS), 5. Glucose Starvation (low glucose media), 6. DTT/PFA (2 mM DTT and 25 mM PFA). Cells were cultured inside T25 flasks at 37 °C, 5% CO₂ for 48 hours, reaching 80-90% confluence. UV radiation: cells were subjected to five cycles of UV irradiation (5 minutes) followed by incubation (5 minutes) to stimulate the production of multivesicular bodies. Exosome isolation: Samples were centrifuged at 500 × g for 5 minutes at 4 °C, and the supernatant was removed. The cell pellet was resuspended in 5 mL PBS and centrifuged again at 952 rpm for 5 minutes at 4 °C. The pellet was discarded, and the supernatant was retained. Ultracentrifugation: The clarified supernatant was transferred to ultracentrifuge tubes and spun at 100,000 g for 7 minutes.