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. Author manuscript; available in PMC: 2018 Mar 7.
Published in final edited form as: Pigment Cell Melanoma Res. 2017 Mar 7;30(2):203–218. doi: 10.1111/pcmr.12564

Figure 1. Mouse melanoma and immortalized melanocytes release exosomes.

Figure 1

Extracellular vesicles were isolated from media conditioned by B16F0 cells (a), Cloudman S91 (b), and Melan-A cells (c) and imaged using SEM (bar indicates 500 nm used for a, b and c). SEM images were representative of more than 3 replicates. A higher magnification is shown as inset in panels (see Figure S1). (d) The diameters of the extracellular vesicles from B16F0 (black solid line, N = 66), Cloudman S91 (black dotted line, N = 62), and Melan-A cells (gray shadowed, N = 123) were estimated from the SEM images using ImageJ and summarized using a density distribution, where the density distributions were offset vertically for clarity. (e) Immunoblotting analysis of common exosome markers, where 20 µg of total protein was loaded in each lane (WCL, whole cell lysate; Exo, exosome lysate). (f) Western blot analysis identified IL12RB2 as being present in B16F0 exosomes, B16F0 cells and 2D6 T cells. GAPDH was a loading control and whole cell lysate from 2D6 T cells was used as a positive control for IL12RB2 expression. (g) The presence of IL12RB2 on the surface of exosomes was detected by flow cytometry using IL-12RB2 mAbs-PE. Unstained exosomes were used as a negative control (gray shaded). Results were representative of three replicates.