Fig. 2
FACS analysis of C16-exo and distribution on iodixanol gradient. a FACS analysis of C16-exo deriving from MNI cells cultivated with medium at different pHs To design the R1 region above instrument background noise only PBS was acquired (upper panel). Note that no events were acquired in this region. (mid panel) example of exosome population. (Lower panel) Histograms representing the green fluorescence intensity distribution of events gated in R1 regions of exosome samples recovered in control, pH 6.7 and pH 6.0 conditions b C16-exo (ctr) (2,5 × 108), C16-exo (pH 6.7) (1,8 × 108), C16-exo (pH 6.0) (2,5 × 108), were loaded at the bottom of iodixanol gradient and subjected to ultracentrifugation for 19 h. For each condition, 2 μl of resulting fractions were FACS counted and each fraction represented as percent of total C16-exo recovered. Fraction densities were determined by refractometry. c An equal volume of each fraction was analyzed for the indicated exosome markers by western blotting. Data shown are representative of three independent experiments