A. miR-5096 transfer from U87 to HMEC, two days after loading. Donor U87, either non-transfected (E) or transfected with mimic (M; 30 nM) or inhibitor (I; 30 nM), were cultured alone for 48 h, then co-cultured with acceptor HMEC (right panel) for 12 h. The miR-5096 levels, relative to U6 snRNA, are means ± SD (*P<0.05 vs empty; Mann-Whitney U test and Kruskal-Wallis test; n = 3). B. Blockage of functional GJIC between U87 and HMEC by miR-5096. Transfected U87 were cultured alone for 48 h, then loaded with calcein and DiL-C18. Labelled U87 (donor) were plated onto HMEC monolayer (acceptor) as described in Fig. 3A. Phase-contrast microphotographs after 5 h of co-culture (representative of 3 experiments; Bar 80 μm). Histogram shows the cell number of HMEC receiving dye (calcein) per U87 (mean ±SD, n=3; **P<0.01 vs control). C. Down-regulation of Cx43 expression in U87 by miR-5096. Immunoblot analysis of Cx43 in whole-cell lysates from transfected U87 (right), cultured alone for 48 h then co-cultured with HMEC for 12h (representative of 3 independent experiments; β-actin as loading control). Histogram shows the miR-5096-mediated down-regulation of Cx43 in U87 and the up-regulation of Cx43 in co-cultured HMEC (mean ± SD; *P<0.05 vs empty; Mann-Whitney U test and Kruskal-Wallis test; n = 3). D. miR-5096 mediated tubulogenesis. Representative micro-photographs of HMEC plated on Matrigel for 5 h with U87 previously loaded with mimic or inhibitor (Bar 80 μm). Histogram showing the number of branch points per field (mean ± SD; **P<0.01 vs empty; Kruskal-Wallis test; n = 3).