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For fluorescence microscopy studies, HL-60 cells and HepG2 cells (seeding densities 1 10 4 cells/well and 8 10 4 cells/well, respectively) were placed in serum-free medium to facilitate their adhesion to coverslips and cultured on lysine-coated 8-well chamber slides (IBIDI) following the procedure detailed by 2.8 Cell surface binding, endocytosis, and downregulation of TfRs For surface binding and endocytosis of AF-labeled proteins, the cells were rinsed twice with ice-cold HEPES-buffered, serum-free media, pH 7.4, and incubated for 1 h on ice with either 500 nM hemeAF546HPX, hemeAF546HPX, or AF488 holo-human Tf

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There are statistics to back that up: In May 2020, researchers from the Perelman School of Medicine at the University of Pennsylvania published a study in the journal JAMA Network Open that found the majority of commercially insured patients who visited the emergency department (ED) for an opioid overdose didnt receive the timely follow-up care known to help prevent a future overdose or death of nearly 6,500 patients treated in EDs nationwide for an overdose or other opioid-related medical complications, only 16 percent accessed opioid use disorder (OUD) medications or another form of treatment within three months of the ED visit. Effective drug and alcohol treatment can help reduce those numbers and keep those who do overdose but survive from becoming a future statistic on a CDC report of annual overdose deaths

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