Multi-component Toxicological Study on Rare Earth Upconversion Nanoparticles

UCNPs@PEG was divided into three groups, 0.0 mg/mL, 0.25 mg/mL, 1.0 mg/mL, and each concentration was repeated for three groups. After co-culture with AML12 cells for 24h, the medium was removed by pipette gun, and sterilized PBS was added for washing 3 times. 300μ L RIPA cell lysate was added to the culture dish. The lysed cells were scraped off with sterile cell scraper, and the lysed cell solution was sucked into the cryopreservation tube.(TandemMassTags) Labeled protein for proteomic study. First extract the total protein in cells, then do concentration determination and SDS-PAGE detection, then use TMT labeled protein after protease hydrolysis, label samples after uniform mixing chromatography separation, and then LC-MS/MS analysis. Chromatographic reaction conditions: A phase H2O-FA (99.9: 0.1, v/v), Phase B ACN-H2O-FA (80: 19.9: 0.1, v/v/v), mass resolution 60000, scan charge-to-mass ratio range 350-1500, collision energy 32, resolution 45000, automatic gain control 2e5. Functional annotation analysis of identified proteins was carried out using common protein databases. Differential proteins were screened and GO, KEGG and interaction analysis were carried out respectively. Cluster thermomap and Veen analysis were used to analyze differential data.