Figure 3.

Exosomic miR-155-mediated cross-talk between human monocytes and neuroblastoma (NBL) cells. A) Quantitative real-time polymerase chain reaction (qRT-PCR) for miR-155 in monocytes cocultured with SK-N-BE(2) or CHLA-255 cells for the indicated time periods. B) qRT-PCR for miR-155 in human monocytes cultured for the indicated time periods with the supernatant (SN) of SK-N-BE(2) cells (normal SN) or with exosome-depleted by ultracentrifugation (depleted SN) of SK-N-BE(2) cells. C) qRT-PCR for miR-155 in human monocytes treated for the indicated time periods with whole exosomes fraction (WEF) isolated from SK-N-BE(2) cells compared with PBS. D) qRT-PCR for miR-155 in human monocytes transfected with total RNA extracted from exosomes of SK-N-BE(2) cells (compared with a total scrambled RNA) for the indicated time periods. E) qRT-PCR for miR-155 in human monocytes transfected with Dotap-miR-21 (miR-21) or Dotap-scrambled RNA (scr) for the indicated time periods. F) qRT-PCR for miR-155 in human monocytes transfected for 48 hours with Dotap-Scrambled (scr group) or Dotap-miR-21 (miR-21 group) alone (left histograms) or in human monocytes transfected for 48 hours with anti-TLR8 siRNA and Dotap-Scrambled (TLR8 siRNA group) or with anti-TLR8 siRNA and Dotap-miR-21 (miR-21+TLR8 siRNA group) (right histograms). G) qRT-PCR for miR-155 in exosomes derived from human monocytes alone (control) or cocultured with SK-N-BE(2) or CHLA-255 cells for the indicated time periods. H) qRT-PCR for miR-155 in SK-N-BE(2) or CHLA-255 cells cocultured with human monocytes for the indicated time periods. I) qRT-PCR for miR-155 in SK-N-BE(2) cells cocultured with human monocytes and in presence of Ruxolitinib (Rux), GW4869, or DMSO (as a control) for the indicated time periods. Relative levels of miR-155 expression were normalized to U6 snRNA. Data are presented as mean ± SD of experiments conducted in triplicate. *P = .003, **P = .002, ***P = .001, ****P < .001, Student’s t test, two-sided.