FIGURE 2.

Dynamics of Bodipy C16 uptake and transformation into phospholipids. (a) Mel501 cells were treated with 7 µM Bodipy C16 for the indicated times or (b) treated for 15 min, washed with H‐BSA and chased in complete medium. Cell associated fluorescence was measured by FC. Data are expressed as mean  ±  SEM (n = 3). (c) Total lipids from cells pulsed with Bodipy C16 for 15 min and chased for different times were extracted and analyzed by HPTLC. The amount of lipid spotted per lane was equivalent to 5 × 10⁴ cells. (d) Lipids from 2 × 10⁴ cells pulsed with Bodipy C16 for 5 h and chased for 2 h and 24 h and Bodipy sEV isolated from the conditioned medium were extracted and analyzed by HPTLC for neutral lipids and (e) phospholipids. The amount of lipid spotted per lane was equivalent to 4 × 10⁴ cells and 8,6 × 10⁸ sEV. Asterisk indicates the sample origin line. Lipids were identified based on comparison to the respective lipid standards. Neutral lipids: TAG, triacylglycerols; DAG, diacylglycerol; chol, cholesterol; C16, Bodipy C16; PL, Polar Lipids. Phospholipids: Cer, ceramide; LBPA, Lysobisphosphatidic acid; CL, cardiolipin; PE, phosphatidylethanolamine; PI, phosphatidylinositol; PS, phosphatidylserine; PC phosphatidylcholine; SM, sphingomyelin.