MicroRNA-29b Enhances Osteoclast Survival by Targeting BCL-2-Modifying Factor after Lipopolysaccharide Stimulation

Figure 2

Overexpression of miR-29b increases the number of OCs by increasing the survival rate. BMMs were prepared and incubated with M-CSF (30 ng/ml) and RANKL (40 ng/ml) for 40 h and washed thoroughly. (a, b) Cells were transfected with 30 nM of miR-29b mimic or con mimic in the presence of M-CSF (30 ng/ml) with or without LPS (50 ng/ml). After the indicated times, total RNA was analyzed by qPCR to quantify the expression of miR-29b. Expression levels with con mimic treatment were set at 1 (a). Cells were incubated for 24 h to quantify the expression of TRAP, calcitonin receptor, ATP6v0d2, and DC-STAMP and for 48 h to count TRAP-positive MNCs per well (b). (c) Cells were transfected with miR-29b mimic or con mimic in the presence of M-CSF and LPS. Cells were washed with medium and analyzed after 6 h to measure TRAP-positive MNCs and annexin V-positive cells. (d, e) Cells were transfected with 30 nM of anti-miR-29b or con inh in the presence of LPS (50 ng/ml) and M-CSF (30 ng/ml). After the indicated times, total RNA was analyzed by qPCR to quantify the expression of miR-29b. Expression levels with the con inh treatment were set at 100 (d). Cells were washed with medium, stimulated with LPS for 6 h, and analyzed to measure TRAP-positive MNCs and annexin V-positive cells. Representative photos of OCs. Scale bar: 200 μm. The Ct values of the genes were within 17.82 and 28.65 cycles. ; ; compared with each corresponding control. Similar results were obtained in three independent experiments.

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