GSM8782048: Co-incubation of OCY-EV and antagomir-NC 1 followed by ultrafiltration, then hepatocytes under FFA conditions; Mus musculus; RNA-Seq
1. Cell Lysis: Disrupt cells using a lysis buffer containing chaotropic salts and β-mercaptoethanol to denature proteins and inhibit RNases. 2. Homogenization: Homogenize the sample to ensure complete cell disruption and RNA release. 3. Phase Separation: Add chloroform and centrifuge to separate RNA into the aqueous phase, while DNA and proteins remain in the organic phase or interphase. 4. RNA Precipitation: Mix the aqueous phase with isopropanol to precipitate RNA, then centrifuge to pellet the RNA. 5. Wash: Wash the RNA pellet with 75% ethanol to remove impurities, followed by centrifugation. 6. Resuspension: Air-dry the pellet and dissolve RNA in RNase-free water or buffer. Vehicle Group: Cells treated with PBS; OCY-EV+ANTA-NC Group: Aged osteocyte extracellular vesicles were incubated with antagomir-NC for 30 minutes followed by ultracentrifugation, with a final extracellular vesicles intervention concentration of 2 μg/ml; OCY-EV+ANTA Group: Aged osteocyte extracellular vesicles were incubated with antagomir-483-5p for 30 minutes followed by ultracentrifugation, with a final extracellular vesicles intervention concentration of 2 μg/ml.