Figure S1.
EVs are released in a Rab27- and CD63-dependent manner. (a and b) MDA-MB-231 cells were transfected with siRNAs targeting Rab27a/b, CD63, or nontargeting (NT) siRNA. Cells were incubated for 48 h in glutamine-replete medium, and EVs were isolated by differential centrifugation before analysis by nanoparticle tracking to obtain size and concentration profiles. Values were normalized to cell number and are expressed as the number of particles of a given size, in 0.5-nm increments, per cell. Values represent the mean of n = 4 independent experiments; dotted line and error bars represent SEM. **, P < 0.01; *, P < 0.05; t test with Welch’s correction. Cell lysates and differential centrifugation pellets were also analyzed by Western blotting for the indicated EV markers. Actin was used as a loading control.

EVs are released in a Rab27- and CD63-dependent manner. (a and b) MDA-MB-231 cells were transfected with siRNAs targeting Rab27a/b, CD63, or nontargeting (NT) siRNA. Cells were incubated for 48 h in glutamine-replete medium, and EVs were isolated by differential centrifugation before analysis by nanoparticle tracking to obtain size and concentration profiles. Values were normalized to cell number and are expressed as the number of particles of a given size, in 0.5-nm increments, per cell. Values represent the mean of n = 4 independent experiments; dotted line and error bars represent SEM. **, P < 0.01; *, P < 0.05; t test with Welch’s correction. Cell lysates and differential centrifugation pellets were also analyzed by Western blotting for the indicated EV markers. Actin was used as a loading control.

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