Effect of SL acyl chain structure on the biophysical properties of HEK cells overexpressing different CerSs. Fluorescence anisotropy of NBD-DPPE (A), Rho-DOPE (B), TMA-DPH (C), t-PnA (D), and long lifetime component of t-PnA fluorescence intensity decay (E) in mock- (black), CerS2- (white), and CerS5-transfected (gray) HEK cells. F: Fluorescence anisotropy of t-PnA (•) and NBD-DPPE (○) represented as a function of the fold-increase in VLC-SLs. All experiments were performed at 37°C. The values are the average (±SD) of n ≥ 3; **P < 0.01; ***P < 0.001.