Figure 1.
Correlation between EVs levels and AML disease burden. (A) In vivo imaging of mice xenografted with Molm-14 cells expressing luciferase at 1, 2, and 3 weeks. Images were taken by IVIS (left), and values were normalized to controls to calculate fold change (right). (B) Correlative analysis of tumor burden between human CD45 (hCD45) levels measured by flow cytometer in both the PB and BM in red vs luciferase activity in gray. (C) EV quantification using NanoSight analyzer from Molm-14 xenografted (red) or control (black) mice. Data show relative vesicles abundance over time (left) and correlation analysis at week 3 of PB and BM (red) vs EV abundance (blue). (D) Cryo-TEM image of Molm-14–derived EVs. Scale bar is 100 nm. (E) Workflow for measuring circulating EVs in the PB of mice bearing mGFP+ AML cells. (F) EV counts per unit volume (1 × 10−4 μL) correspond to BM tumor burden. (G) Quantification of relative AML EV count in the PB compared with total lipid vesicles in plasma. Each point represents EV count per unit volume (1 × 10−4 μL). (H) Representative 3-dimensional images and concentration of 20% (top) and 50% (bottom) animal cohorts. Bounding box represents 100 μm × 100 μm × 10 μm volume. Concentrations determined using [EVs/µL = (average EV count) × (volume/1) × (dilution factor)]. mGFP (white). (I) PB plasma of AML xenografted NSG mice contain numerous lipid vesicles and a discrete mGFP EV population (right) not seen in nonengrafted control animals (left). CellMask Lipid Dye (red), mGFP (green). Bounding box is 5 μm x 5 μm. Images captured with CoreDV epifluorescence microscope with 100× 1.49 TIRF objective and Nikon Coolpix CCD camera. Significance determined by Student t test. *P < .05; ***P < .001.

Correlation between EVs levels and AML disease burden. (A) In vivo imaging of mice xenografted with Molm-14 cells expressing luciferase at 1, 2, and 3 weeks. Images were taken by IVIS (left), and values were normalized to controls to calculate fold change (right). (B) Correlative analysis of tumor burden between human CD45 (hCD45) levels measured by flow cytometer in both the PB and BM in red vs luciferase activity in gray. (C) EV quantification using NanoSight analyzer from Molm-14 xenografted (red) or control (black) mice. Data show relative vesicles abundance over time (left) and correlation analysis at week 3 of PB and BM (red) vs EV abundance (blue). (D) Cryo-TEM image of Molm-14–derived EVs. Scale bar is 100 nm. (E) Workflow for measuring circulating EVs in the PB of mice bearing mGFP+ AML cells. (F) EV counts per unit volume (1 × 10−4 μL) correspond to BM tumor burden. (G) Quantification of relative AML EV count in the PB compared with total lipid vesicles in plasma. Each point represents EV count per unit volume (1 × 10−4 μL). (H) Representative 3-dimensional images and concentration of 20% (top) and 50% (bottom) animal cohorts. Bounding box represents 100 μm × 100 μm × 10 μm volume. Concentrations determined using [EVs/µL = (average EV count) × (volume/1) × (dilution factor)]. mGFP (white). (I) PB plasma of AML xenografted NSG mice contain numerous lipid vesicles and a discrete mGFP EV population (right) not seen in nonengrafted control animals (left). CellMask Lipid Dye (red), mGFP (green). Bounding box is 5 μm x 5 μm. Images captured with CoreDV epifluorescence microscope with 100× 1.49 TIRF objective and Nikon Coolpix CCD camera. Significance determined by Student t test. *P < .05; ***P < .001.

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