Figure 4.

Differentially Expressed miRNAs in OLF and the Function of miR-19b-3p

(A) Sequencing analysis for miRNA was performed with RNA extracted from ossification (n = 4) and normal (n = 4) LF. Hierarchical cluster analysis of significantly differentially expressed lncRNAs: bright green, under-expression; gray, no change; bright red, overexpression. (B) Differential expression of ten representative miRNAs was validated in human ossified and normal LF tissues by qPCR (n = 10 per group). (C) The differentially expressed miRNAs were distributed on different chromosomes. (D and E) qPCR analysis of miR-19b-3p was performed with total RNA isolated from ossified and normal LF (D) and hMSCs treated with osteogenic medium for 0 and 7 days (E). (F) hMSCs were transfected with miR-19b-3p mimic or the negative control and further cultured in osteogenic medium for 7 days. The RNA levels of miR-19b-3p, ALP, Col1α1, BGLAP, and RUNX2 were detected by qPCR. U6 was used as an internal control for miR-19b-3p. GAPDH was used as an internal control for mRNA. (G) Representative images of ALP staining of hMSCs after transfection with miR-19b-3p mimic or the negative controls and further culture in osteogenic medium for 7 days. Scale bars, 400 μm. Data are presented as mean ± SEM for at least triplicate experiments. The p values were analyzed by Student’s t test; *p < 0.05.