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. 2015 Jun 18;6:7321. doi: 10.1038/ncomms8321

Figure 2. Functional transfer of miR-155 via exosomes in vitro.

Figure 2

(a) Schematic of the exosome transfer experiment. (b) qRT–PCR was used to measure relative miR-155 levels in miR-155−/− BMDCs that received either Wt or miR-155−/− exosomes derived from BMDCs treated with or without GW4869 (n=5). (c,d) mRNA levels of miR-155 targets, BACH1 and SHIP1, from the same experiment shown in b as measured by qRT–PCR (n=5). (e) Representative western blottings of SHIP1 and β-actin in miR-155−/− BMDCs given either Wt or miR-155−/− exosomes. (f) Protein levels of SHIP1 were quantified using ImageJ software (n=2). (g) Relative miR-155 levels in Wt BMDCs given either Wt or miR-155−/− exosomes as quantified by qRT–PCR (n=6). (h,i) BACH1 and SHIP1 mRNA levels were measured in the same experiment shown in g as quantified by qRT–PCR (n=6). (j) qRT–PCR was used to quantify HO1 mRNA levels during the experiment in b (n=5). (k) Western blotting for AGO2 and β-actin from miR-155−/− BMDCs given Wt or miR-155−/− exosomes. On the left is the input (whole-cell lysate), the middle is from the pan-AGO pulldown where one-third of input was used and the right is the IgG pulldown where one-third of the input was used. (l) Relative miR-155 levels were quantified via qRT–PCR in the same experiment shown in k. (m) miR-146a levels were quantified using qRT–PCR during the experiment in k. Levels in l,m are plotted as Ago:IgG. Dotted line separates input from pull-down groups. Data represent two independent experiments and are presented as the mean±s.d. (error bars). *P<0.05; **P<0.01, Student's t-test.