Figure 3.

(Overleaf.) EVs secreted by MYCN-expressing cells promote glycolysis, respiration, ATP production and proliferation of recipient neuroblastoma cells. (a) Fluorescence microscopy of SH-EP cells after 24 h incubation with PKH67-labelled EVs isolated from TET21-N cells expressing or not expressing MYCN. Quantification of the percentages of cells positive to the dye in different conditions is shown on (iii). Error bars indicate mean values ± s.e.m. Scale bar 25 μm. (b) SH-EP cells were counted at 1 day intervals after being cultured in the presence or absence of EVs purified from MYCN-positive or -negative TET21-N cells (n = 3). (c) SH-EP cells were subjected to MTS assays 24 or 48 h after being cultured in the presence or absence of EVs purified from MYCN-positive or -negative TET21-N cells (n = 5). (d) L-lactate production in SH-EP (n = 4) and (e) SH-SY5Y (n = 3) neuroblastoma cells incubated for 24 h with EVs isolated from TET21-N cells with and without MYCN expression. (f–j) Seahorse analysis. MYCN single-copy cell line SH-SY5Y was used as a recipient for EVs purified from TET21-N cells with or without MYCN (n = 3). PBS was used as a control (untreated). OCR indicates oxygen consumption rate. Error bars represent mean values ± s.e.m. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001.