Fig. 3.

MCF-7 cell-derived miR-375 is taken up by MΦ via CD36. a MCF-7 cell ACM was prepared with (+ FCS) or without (− FCS) FCS, and miR-375 and miR-183-5p abundance was measured by qPCR. Synthetic cel-miR-39a was used as spike-in control for RNA purification efficiency and normalization control (n = 4). b LDL and HDL fractions were separated from ACM (with or without FCS in the media) and analyzed for the abundance of miR-375 and miR-183-5p by qPCR (n = 3). c, d MΦ were pre-incubated with IgG control or anti-CD36 blocking mAb for 1 h and during the whole experiment. c MΦ were treated with ACM for 30 min. Cells were washed and further cultured in MΦ media for 24 h. MiR-375 abundance was measured by qPCR and normalized to untreated control MΦ (n = 7). d MΦ were cocultured with MCF-7 cells for 24 h and miR-375 abundance was measured via qPCR, and normalized to untreated control MΦ (n = 4). e MΦ were treated with MCF-7 ACM alone or together with a CD36-blocking peptide for 30 min. The peptide was added 1 h before ACM treatment. Afterwards, cells were washed and further cultured in MΦ media alone or together with the peptide for 24 h. miR-375 level was measured by qPCR and normalized to untreated control MΦ (n = 8). f, g MΦ were transfected with control (nonspecific siRNA) or CD36 siRNA for 24 h and cocultured with MCF-7 cells for another 24 h. f CD36 mRNA expression and g miR-375 levels were measured by qPCR (n = 6). Data of a–g are mean ± SEM and p-values were calculated using two-tailed Student’s t-test (a, b, f, g) and one-sample t-test (c–e); *p < 0.05, **p < 0.01, n.s., not significant