Figure 4
From: LAMP3 transfer via extracellular particles induces apoptosis in Sjögren’s disease
Extracellular particles from LAMP3-overexpressing cells contain LAMP3. (A) Western blot analysis of EPs separated from the culture medium of control, LAMP3-overexpressing, and LAMP1-overexpressing A253 cells using centrifugation in combination with or without total exosome isolation reagent. Equal volume of protein loaded in each lane. Lower panel shows same membrane as the one used in Western blot analysis but stained with Reversible Protein Stain Kit. Uncropped images are provided in Supplementary Fig. 3. (B) Cell surface LAMP3 expression was analyzed by flow cytometry (bold line: LAMP3 staining, filled area with dashed line: isotype control) and under an immunofluorescent microscopy (scale bar: 100 μm) in control, LAMP1-overexpressing, and LAMP3-overexpressing A253 cells 48 h post-transfection. (C) EPs derived from control and LAMP3-overexpressing A253 cells were immunoprecipitated using anti-LAMP3 monoclonal antibody-conjugated beads. Presence of LAMP3 protein on EP membrane surface was determined by analyzing beads stained with anti-LAMP3 polyclonal antibody using flow cytometry. Graph showing difference in mean (± SD) percentage of LAMP3-positive beads compared with beads bound to isotype control (n = 4). **p < 0.01 (one-way ANOVA).
