Representative confocal microscopy images (top) of C6-ceramide uptake and quantification (bottom) of fluorescent spots resembling intracellular vesicles containing the corresponding lipid probe. The uptake was assayed on HeLa cells following transient expression of the WT, p.Ser800Leu, p.Asp849Glu, and p.His852Tyr DENND5B constructs. Quantification of spots was performed on TagBFP2-positive cells. Data are means ± SEM (n = 5). For each independent biological replicate, a region measuring 1,320 x 1,320 μm was imaged and analyzed, comprising approximately 35–40 TagBFP2-positive cells.
