After 4 h of treatment, cells were detached using the cell scraper and 2,5 10 5 cells were seeded in the upper chamber of Transwell chambers equipped with 5-m pore-size polycarbonate membranes, as already described (Pijuan et al., 2019)
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Isolated CD4 + and CD8 + T cells were activated or not for up to 1, 3, or 6 days in a reverse time course with Dynabeads (1:1 cell-to-bead ratio, Gibco), magnetic beads similar in size to antigen-presenting cells and coupled with anti-CD3 and anti-CD28 antibodies, so that their total time in culture would be identical and to ensure that all cells would finish their indicated activation period the same day
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State Legislative Landscape in 2025 Despite a significant number of bills in 2023 and 2024 that would have required mandated coverage of GLP-1s in Medicaid, state employee health plans, or for health insurers regulated by the state, only Arkansas, California, Maryland, New Mexico, Pennsylvania, and Texas have introduced legislation mandating coverage in 2025 to date
