Renal tubular VMP1 protects against acute kidney injury via facilitating autophagic flux

Acute kidney injury (AKI) is intricately linked to autophagy. Our study investigates the pivotal role of the autophagy-associated protein VMP1 in the pathogenesis and progression of AKI. Overall design: 48 hours after HK2 cells were transfected with FLAG-VMP1 plasmid, RIP (RNA immunoprecipitation) was performed using Flag antibody magnetic beads, and the extracted RNA was sequenced by transcriptome.The protocol of RIP:To perform an RNA Binding Protein Immunoprecipitation (RIP) assay, first lyse cells in a buffer containing protease and RNase inhibitors. Pre-clear the lysate with protein A or G beads, then add specific antibodies against the target RNA-binding protein and incubate overnight at 4°C. After incubation, add the beads again and continue to incubate for several hours to allow binding. Wash the beads extensively to remove non-specific interactions, and then elute the bound RNA. Finally, extract and analyze the RNA using methods such as RT-qPCR or RNA sequencing to identify the associated RNA species.