Fig. 10

Silencing CD133 affects the distribution of lipid droplets and impedes the biogenesis of extracellular lipidosomes. A-D Native or CD133-deficient (clone –/5) FEMX-I cells growing either on poly-L-lysine (A) or fibronectin-coated (A-D) supports were either processed for CLSM (A, B) or recorded in live by phase-contrast/fluorescence video microscopy after staining with LipidSpot™ 610 (C, D). For CLSM, PFA-fixed cells without (A) or with (B) saponin-permeabilization were co-stained with BODIPY™ 493/503 and fluorescence-conjugated WGA (A) to highlight lipid droplets and glycoconjugates at the cell membrane, respectively, or double-immunolabeled with antibodies directed against CD133 (293C3) and adipophilin (B). For the live cell imaging, elapsed time in minutes is shown on the top-right corner. The images are excerpted from the Additional file 11: Video S10 (C) and Additional file 12: Video S11 (D). Note the EVs lacking lipid droplets during cell division of CD133-deficient cells (D, arrowhead). Asterisks indicate the concentration of lipid droplets at cell extremities of native cells (A, B), while arrows indicate the retraction of cell extremities (D). The dashed lines mark the outline of the CD133-deficient cells (B). Scale bars are indicated