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. 2020 Dec 23;8:594785. doi: 10.3389/fcell.2020.594785

FIGURE 4.

FIGURE 4

MiR-1247-5p is a complementary targeting miRNA of circHmbox1. (A) The expressions of miRNAs binding to circHmbox1 were analyzed by qRT-PCR in TNF-α-induced osteoclasts differentiation. n = 3, **P < 0.01. (B) Complementation between circHmbox1 and “seed sequence” of miR-1247-5p were showed with yellow highlights. (C) Dual luciferase reporter assay displayed the complementary binding of circHmbox1 and miR-1247-5p. n = 6, **P < 0.01. (D) The expressions of miR-1247-5p were analyzed by qRT-PCR in osteoclasts overexpressing or silencing miR-1247-5p. n = 3, **P < 0.01. (E) The expressions of miR-1247-5p were analyzed by qRT-PCR in osteoclasts overexpressing or silencing miR-1247-5p in the presence of TNF-α. n = 3, **P < 0.01. (F) TRAP staining was accomplished in osteoclasts overexpressing/inhibiting miR-1247-5p in the presence of RANKL and M-CSF for 5 days. (G) Summarized data showed that miR-1247-5p could promote RANKL-induced osteoclast differentiation. However, inhibition of miR-1247-5p could alleviate the induction of RANKL on osteoclast differentiation. n = 4, **P < 0.01. (H) TRAP staining was accomplished in osteoclasts overexpressing/inhibiting miR-1247-5p in the presence of RANKL, M-CSF and TNF-α for 5 days. (I) Summarized data showed that miR-1247-5p promoted TNF-α-induced osteoclast differentiation. AMO-1247-5p attenuated the promoting effect of TNF-α on osteoclasts differentiation. n = 4, **P < 0.01.