Figure 6. Osteoclast-derived exosomal miR-214-3p transfer to osteoblasts.

(a) A schematic diagram illustrating the design of co-culture experiments with GFP-exosome-producing osteoclasts and osteoblasts. CMV-GFP-CD63 were transfected into the OC-miR-214-3p osteoclasts to label the osteoclast-derived exosomes. (b) Representative confocal images of the GFP-exosome-producing osteoclasts at 24 h after transfection (left panels), and the uptake of GFP-exosome by osteoblasts (right panels) at 24 h after co-culture. (c) A schematic diagram illustrating the design of co-culture experiments with the osteoblasts and miR-214-3p-depleted osteoclasts (upper panels), and real-time PCR analysis of the levels of pri-miR-214-3p, pre-miR-214-3p and miR-214-3p in osteoblasts (normalized by the mean value of No OC group) at 24 h after co-culture with miR-214-3p-depleted/intact osteoclasts differentiated from miR-214-3p-depleted/intact RAW264.7 cells, respectively (lower panels). (d) A schematic diagram illustrating the design of co-culture experiments with the osteoclasts and miR-214-3p-depleted osteoblasts (upper panels), and real-time PCR analysis of the miR-214-3p level in miR-214-3p-depleted osteoblasts (normalized by the mean value of No OC group) at 24 h after co-culture with either OC-miR-214-3p or WT osteoclasts (lower panels). All data are the mean±s.d. of four independent experiments. *P<0.05. One-way ANOVA with a post-hoc test was performed.