Journal: Oncotarget

Article Title: Exosomal survivin facilitates vesicle internalization

doi: 10.18632/oncotarget.26182

Figure Lengend Snippet: Exosomal surface proteins were digested by proteinase K (“shaving”) and then introduced to cell culture. (A) Untreated and treated HeLa cells fixed and imaged with fluorescent microscopy (original magnification 40x; blue, DAPI stained nuclei; green, PKH67 stained exosomes.) Proteinase K treatment did not affect the PKH67 staining of exosomal membranes. (B) Cell count using the fluorescent imaging as described above. Cells staining green were counted as PKH67 positive cells, showing a higher percentage of internalization of intact exosomes than “shaved” exosomes. Total cell number was comparative between groups. (C) PKH67 labelled “shaved” exosomes were internalized at a lower rate than intact exosomes as shown by flow cytometry.

Article Snippet: Phase contrast images with fluorescent PKH67 stained exosomes were imaged on the Keyence BZ-X700 all-in-one fluorescent microscope (Keyence Corporation, Itasca, IL).

Techniques: Cell Culture, Microscopy, Staining, Cell Counting, Imaging, Flow Cytometry

Journal: Oncotarget

Article Title: Exosomal survivin facilitates vesicle internalization

doi: 10.18632/oncotarget.26182

Figure Lengend Snippet: HeLa cells co-incubated with PKH67 stained exosomes and soluble Survivin show a significant reduction in exosome uptake in a dose dependent manner. Data is representative of 3 independent experiments. Significance was determined by one-way ANOVA with ad hoc Tukey’s multiple comparison’s tests * p<0.05, ** p<0.01, *** p<0.005, **** p<0.001.

Article Snippet: Phase contrast images with fluorescent PKH67 stained exosomes were imaged on the Keyence BZ-X700 all-in-one fluorescent microscope (Keyence Corporation, Itasca, IL).

Techniques: Incubation, Staining

Journal: Oncotarget

Article Title: Exosomal survivin facilitates vesicle internalization

doi: 10.18632/oncotarget.26182

Figure Lengend Snippet: As a control, cells were incubated with an antibody to clathrin, an intracellular protein involved in endocytosis. As expected due to the inability of the antibody to cross the membrane barrier, no change was measured in the uptake of PKH67 stained exosomes by flow cytometry. Data is representative of 3 or more experiments. Experiments were analyzed on flow cytometry and data is presented with median fluorescence intensity (MFI). Significance was determined by student’s t-test or one-way ANOVA with ad hoc Tukey’s multiple comparison’s tests * p<0.05, ** p<0.01, *** p<0.005, **** p<0.001.

Article Snippet: Phase contrast images with fluorescent PKH67 stained exosomes were imaged on the Keyence BZ-X700 all-in-one fluorescent microscope (Keyence Corporation, Itasca, IL).

Techniques: Control, Incubation, Membrane, Staining, Flow Cytometry, Fluorescence

Journal: Oncotarget

Article Title: Exosomal survivin facilitates vesicle internalization

doi: 10.18632/oncotarget.26182

Figure Lengend Snippet: Reduction of PKH67 stained exosome internalization following antibody blocking of transferrin receptor 1 (TfR1), glucocorticoid receptor (GR), endothelin B receptor (ETBR), and insulin receptor alpha (IRα). Data is representative of 3 or more experiments. Experiments were analyzed on flow cytometry and data is presented with median fluorescence intensity (MFI). Significance was determined by student’s t-test or one-way ANOVA with ad hoc Tukey’s multiple comparison’s tests * p<0.05, ** p<0.01, *** p<0.005, **** p<0.001.

Article Snippet: Phase contrast images with fluorescent PKH67 stained exosomes were imaged on the Keyence BZ-X700 all-in-one fluorescent microscope (Keyence Corporation, Itasca, IL).

Techniques: Staining, Blocking Assay, Flow Cytometry, Fluorescence

Journal: Oncotarget

Article Title: Exosomal survivin facilitates vesicle internalization

doi: 10.18632/oncotarget.26182

Figure Lengend Snippet: No change in PKH67 stained exosome internalization following antibody blocking of TfR2, tumor necrosis factor receptors (TNFR1 and TNFR2), and low-density lipoprotein receptor (LDLR). Data is representative of 3 or more experiments. Experiments were analyzed on flow cytometry and data is presented with median fluorescence intensity (MFI). Significance was determined by student’s t-test or one-way ANOVA with ad hoc Tukey’s multiple comparison’s tests * p<0.05, ** p<0.01, *** p<0.005, **** p<0.001.

Article Snippet: Phase contrast images with fluorescent PKH67 stained exosomes were imaged on the Keyence BZ-X700 all-in-one fluorescent microscope (Keyence Corporation, Itasca, IL).

Techniques: Staining, Blocking Assay, Flow Cytometry, Fluorescence

Journal: Oncotarget

Article Title: Exosomal survivin facilitates vesicle internalization

doi: 10.18632/oncotarget.26182

Figure Lengend Snippet: (A) HeLa cells pre-treated with antibodies to Survivin showed no change in uptake of PKH67 stained exosomes as assessed with flow cytometry. (B) PKH67 stained exosomes pre-incubated with Survivin antibody show a decreased amount of internalization by HeLa cells. Data is representative of 3 independent experiments. Significance was determined by one-way ANOVA with ad hoc Tukey’s multiple comparison’s tests * p<0.05, ** p<0.01, *** p<0.005 , **** p<0.001.

Article Snippet: Phase contrast images with fluorescent PKH67 stained exosomes were imaged on the Keyence BZ-X700 all-in-one fluorescent microscope (Keyence Corporation, Itasca, IL).

Techniques: Staining, Flow Cytometry, Incubation