Fig. 4
Inhibitor of differentiation 4 (ID4) expression in breast cancer cells leads to the paracrine downregulation of miR-107 in macrophages. a and b RT-qPCR analysis to evaluate the expression of miR-107 in macrophages obtained from 1,25-dihydroxyvitamin D3 (VitD3)-mediated differentiation of HL60 (a) and U937 (b) cells and subsequently cultivated in conditioned media (CM) from control (si-SCR) or ID4-depleted (si-ID4) MDA-MB-468 cells. c RT-qPCR for miR-96 in U937-derived macrophages as in (b). d and e RT-qPCR analysis of miR-107 in peripheral blood-derived macrophages (PBD-M) (d) and U937-derived (e) macrophages cultivated in RPMI medium (CTR) or in CM from, respectively, SKBR3 and MDA-MB-468 cells for the indicated time points. f RT-qPCR for pre-miR-107 in U937 cells as in (e). g RT-qPCR analysis of miR-107 levels in differentiated U937 cells transfected with locked nucleic acid (LNA) antisense oligonucleotide directed to miR-107. h Western blot analysis of the indicated proteins in differentiated U937 cells transfected with LNA antisense oligonucleotide directed to miR-107. i and j miR-107 (i) and granulin (GRN) (j) expression levels evaluated by RT-qPCR in HL60 and U937 cells transfected with control mimic or miR-107 mimic oligonucleotides. k Western blot analysis of GRN in HL60 and U937 cells transfected with control mimic or miR-107 mimic oligonucleotides. Results from at least three biological replicates are shown. Data are presented as mean ± SEM. *P < 0.05, **P < 0.005, ***P < 0.0005 calculated by two-tailed t test.