Quantification of Neutral Lipids in Treated BUVEC

BUVEC (n = 5) were seeded into 25 cm² flasks (Sarstedt) and cultured until confluence. Then, the cells were treated with BLT-1 (2 µM) for 48 h. To determine if inhibitor treatments exerted an effect on neutral lipids, pre-treated cells were stained with BODIPY 493/503 (2 µg/mL, 1 h, 37 °C, 5% CO₂, Cayman Chemical). Afterwards, cells were washed twice in 1× PBS (600× g; 5 min) and samples were analyzed with a BD Accuri C6^® FACS cell analyzer (Becton-Dickinson, Heidelberg, Germany). The cells were gated according to their size and granularity, while BODIPY 493/503-derived signals were assessed in the FL-1 channel as described elsewhere [15 (link)].