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. 2017 Jun 16;113(6):1342–1352. doi: 10.1016/j.bpj.2017.05.032

Figure 5.

Figure 5

Characteristics of serum-triggered EV release. (A and B) HEK293 cells grown for 16 h without serum were incubated in fresh medium without (−) or with (+) 10% serum for 4 h. EVs harvested by PEG/ConA were immunoblotted for CD63 and CD9 and quantitated across three experiments, as shown. (C) HEK293 cells grown for 16 h without serum were incubated in fresh serum-free media for 1 h (0). This was collected and replaced with fresh medium containing 10% serum every hour for 4 h (1–4). EVs were harvested by PEG/ConA precipitation and immunoblotted for CD63 and CD9. (D) Serum-triggered EVs from control and VPS4EQ-expressing cells were harvested by PEG/ConA and immunoblotted for CD63 and CD9. (E) Quantitation of three independent experiments. Values show the percent of protein in PEG/ConA EVs from serum stimulated cells expressing VPS4AEQ compared to EVs from control serum stimulated cells not induced to express VPS4AEQ. The lines indicate mean.