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. 2016 Mar 7;7:10872. doi: 10.1038/ncomms10872

Figure 5. Osteoclastic miR-214-3p inhibit osteoblast activity.

Figure 5

(a) A schematic diagram illustrating the design of the co-culture experiments. The osteoblasts derived from the calvarial bone of newborn C57BL/6 mice were co-cultured with the osteoclasts derived from OC-miR-214-3p mice (OC-miR-214-3p OCs) or WT mice (WT OCs), or cultured without osteoclasts (No OC). (b) Real-time PCR analysis of the supernatant, supernatant exosomal and intra-osteoblast miR-214-3p levels (normalized by the mean value of No OC) at 24 h after co-culture, respectively. (c) The pri-miR-214-3p and pre-miR-214-3p levels in osteoblasts at 24 h after co-culture. (d) Real-time PCR analysis of the mRNA levels of osteoblast activity-related marker genes (Alp, Opn, BSP and Bglap) in osteoblasts at 48 h after co-culture. (e) Real-time PCR analysis of the mRNA expression levels of Alp, Opn, BSP and Bglap in osteoblasts transfected with exogenous ATF4 mRNA 3′UTR at 48 h after co-culture. The osteoblasts transfected with either vehicle (OC-miR-214 3p+Veh), nonsense 3′UTR control (OC-miR-214 3p+NC) or ATF4 mRNA 3′UTR (OC-miR-214 3p+ ATF4 mRNA 3′UTR) were co-cultured with the OC-miR-214 3p osteoclasts. All data are the mean±s.d. of four independent experiments. *P<0.05. One-way ANOVA with a post-hoc test was performed.