Fig. 3

Human LGALS3BP is overexpressed and secreted by engineered murine neuroblastoma NXS2 hLGALS3BP cells. (A) Histograms showing transcriptional levels of human (hLGALS3BP) and murine (mLGALS3BP) LGALS3BP in NXS2 CTRL and hLGALS3BP cells normalized to housekeeping gene (β-actin). Values refer to two independent biological replicates. (B) Western blotting images showing intracellular protein levels of hLGALS3BP in NXS2 CTRL and hLGALS3BP cells. Equal amounts of protein were loaded for each sample. β-actin was used as loading control. Images are representative of two independent biological replicates. (C) Secreted levels (ng/mL) of hLGALS3BP and mLGALS3BP assessed by ELISA assay in NXS2 CTRL and hLGALS3BP cells. Values refer to two independent biological replicates. (D) Table showing secreted LGALS3BP expression level in a panel of human cancer cell lines. (E) Confocal imaging of NXS2 CTRL and hLGALS3BP cells showing LGALS3BP (green) and nuclei (Hoechst). Scale bar: 10 μm. (F) Immunoblot showing EVs-associated hLGALS3BP and positive/negative exosomal markers expression in whole cell lysates (WCL) or EVs isolated from NXS2 CTRL or hLGALS3BP. Equal amount of protein were loaded for each sample.