Additional file 6: of Acidic microenvironment plays a key role in human melanoma progression through a sustained exosome mediated transfer of clinically relevant metastatic molecules

Figure S5. MNI cells labeling with CFDA-SE, vesicles isolation and FACS analysis. a MNI cells were left untreated, or pH 6.7, and pH 6.0 treated for 22 h followed by 0.3 μM CFDA-SE labeling 5 min at room temperature, then FACS analyzed. Bars: mean ± S.D. (n = 3). b Workflow of vesicles isolation from MNI cells. Vesicles were labelled with (10 μM) CFDA-SE for 1 h at room temperature to obtain CFSE-EV. c FACS analysis of CFSE-EV population deriving from MNI cells cultivated at different pHs. To design the R1 region above instrument background noise only PBS was acquired. Note that no events were acquired in this region. Histograms represent the green fluorescence intensity distribution of events gated in R1 regions of CFSE population recovered from MNI cells in standard culture condition (ctr), pH 6.7 and pH 6.0. (PNG 383 kb