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OB uptake of T47D EVs is highest at 24 h. (A) Workflow schematic illustrating OB exposure to T47D-EV preconditioning. (B) Representative NTA histogram of concentration and size distribution of EVs remaining in the culture media of T47D EV-treated OBs at 0, 6, 12, and 24 h (n = 3). (C) Representative NTA histogram of concentration and size distribution of EVs in the culture media of OBs cultured in TBBM at 6, 12, and 24 h (n = 3). (D) Representative images of T47D-EV uptake after 24 h with nuclei of OBs stained with DAPI (blue), the cytoplasm of OBs stained with CFSE (green), and pre-stained T47D EVs internalized within OBs stained with <t>MemGlow</t> (magenta). Scale bar represents 50 µm.
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OB uptake of T47D EVs is highest at 24 h. (A) Workflow schematic illustrating OB exposure to T47D-EV preconditioning. (B) Representative NTA histogram of concentration and size distribution of EVs remaining in the culture media of T47D EV-treated OBs at 0, 6, 12, and 24 h (n = 3). (C) Representative NTA histogram of concentration and size distribution of EVs in the culture media of OBs cultured in TBBM at 6, 12, and 24 h (n = 3). (D) Representative images of T47D-EV uptake after 24 h with nuclei of OBs stained with DAPI (blue), the cytoplasm of OBs stained with CFSE (green), and pre-stained T47D EVs internalized within OBs stained with <t>MemGlow</t> (magenta). Scale bar represents 50 µm.
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OB uptake of T47D EVs is highest at 24 h. (A) Workflow schematic illustrating OB exposure to T47D-EV preconditioning. (B) Representative NTA histogram of concentration and size distribution of EVs remaining in the culture media of T47D EV-treated OBs at 0, 6, 12, and 24 h (n = 3). (C) Representative NTA histogram of concentration and size distribution of EVs in the culture media of OBs cultured in TBBM at 6, 12, and 24 h (n = 3). (D) Representative images of T47D-EV uptake after 24 h with nuclei of OBs stained with DAPI (blue), the cytoplasm of OBs stained with CFSE (green), and pre-stained T47D EVs internalized within OBs stained with <t>MemGlow</t> (magenta). Scale bar represents 50 µm.
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OB uptake of T47D EVs is highest at 24 h. (A) Workflow schematic illustrating OB exposure to T47D-EV preconditioning. (B) Representative NTA histogram of concentration and size distribution of EVs remaining in the culture media of T47D EV-treated OBs at 0, 6, 12, and 24 h (n = 3). (C) Representative NTA histogram of concentration and size distribution of EVs in the culture media of OBs cultured in TBBM at 6, 12, and 24 h (n = 3). (D) Representative images of T47D-EV uptake after 24 h with nuclei of OBs stained with DAPI (blue), the cytoplasm of OBs stained with CFSE (green), and pre-stained T47D EVs internalized within OBs stained with <t>MemGlow</t> (magenta). Scale bar represents 50 µm.
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OB uptake of T47D EVs is highest at 24 h. (A) Workflow schematic illustrating OB exposure to T47D-EV preconditioning. (B) Representative NTA histogram of concentration and size distribution of EVs remaining in the culture media of T47D EV-treated OBs at 0, 6, 12, and 24 h (n = 3). (C) Representative NTA histogram of concentration and size distribution of EVs in the culture media of OBs cultured in TBBM at 6, 12, and 24 h (n = 3). (D) Representative images of T47D-EV uptake after 24 h with nuclei of OBs stained with DAPI (blue), the cytoplasm of OBs stained with CFSE (green), and pre-stained T47D EVs internalized within OBs stained with <t>MemGlow</t> (magenta). Scale bar represents 50 µm.
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OB uptake of T47D EVs is highest at 24 h. (A) Workflow schematic illustrating OB exposure to T47D-EV preconditioning. (B) Representative NTA histogram of concentration and size distribution of EVs remaining in the culture media of T47D EV-treated OBs at 0, 6, 12, and 24 h (n = 3). (C) Representative NTA histogram of concentration and size distribution of EVs in the culture media of OBs cultured in TBBM at 6, 12, and 24 h (n = 3). (D) Representative images of T47D-EV uptake after 24 h with nuclei of OBs stained with DAPI (blue), the cytoplasm of OBs stained with CFSE (green), and pre-stained T47D EVs internalized within OBs stained with MemGlow (magenta). Scale bar represents 50 µm.

Journal: Frontiers in Pharmacology

Article Title: Extracellular vesicle-mediated preconditioning of osteoblasts by ER+ breast cancer cells: implications for the bone metastatic niche

doi: 10.3389/fphar.2026.1844422

Figure Lengend Snippet: OB uptake of T47D EVs is highest at 24 h. (A) Workflow schematic illustrating OB exposure to T47D-EV preconditioning. (B) Representative NTA histogram of concentration and size distribution of EVs remaining in the culture media of T47D EV-treated OBs at 0, 6, 12, and 24 h (n = 3). (C) Representative NTA histogram of concentration and size distribution of EVs in the culture media of OBs cultured in TBBM at 6, 12, and 24 h (n = 3). (D) Representative images of T47D-EV uptake after 24 h with nuclei of OBs stained with DAPI (blue), the cytoplasm of OBs stained with CFSE (green), and pre-stained T47D EVs internalized within OBs stained with MemGlow (magenta). Scale bar represents 50 µm.

Article Snippet: EVs were fluorescently labeled with MemGlow (Cytoskeleton, MG04) at a final concentration of 0.2 μM and incubated for 20 min at room temperature in Protein LoBind tubes (Eppendorf, 0030108116).

Techniques: Concentration Assay, Cell Culture, Staining