Figure 1: osteogenic induction upregulated lnc-RNA H19 expression and osteogenesis related gene expression. (A) alizarin red staining of mineralization core in BMSC cell culture after incubated with osteogenic induction medium for 7 days. Volume of mineralization core was counted and analyzed by statistics. (B) RNA expression of H19 was tested by real time PCR. OST-BMSC stands for osteogenic induction of BMSC cells by 7 days. (C, D, E) Runx2, collagen I and ALP mRNA expression level was examined by real-time PCR after osteogenic induction of BMSCs for 7 days. GAPDH is internal control for equal calculation. *P < 0.05, ***P < 0.01.

Figure 2: Overexpression of lnc-RNA H19 enhanced viability of BMSC cells and induced BMSC osteogenesis differentiation markers. (A) apoptotic cells were examined by flow cytometry after overexpression of H19 for 48 h, Annexin-V+ areas were calculated and compared with control groups. (B) MTT assay was performed after overexpression of H19 in BMSC cells for 24 h and 48 h, respectively. (C, D, E) Runx2, collagen I and ALP mRNA expression level was examined by real-time PCR after overexpression of H19 for 48 h. GAPDH is internal control for equal calculation. *P < 0.05.

Figure 3: Knockdown of lnc-RNA H19 inhibited viability of BMSC cells. Si-NC represented negative control siRNA and has no effect to H19 expression. (A) MTT assay was performed after knockdown of H19 for 24 h and 48 h, respectively. (B, C) Apoptotic cells were counted after knockdown of H19 for 48 h by flow cytometry assay. (D, E, F) Runx2, collagen I and ALP mRNA expression level was examined by real-time PCR after knockdown of H19 for 48 h. GAPDH is internal control for equal calculation. *P < 0.05.

Figure 4: lnc-RNA H19 was either overexpressed or knocked down for 48 h. Expression of BMP2 (A), Wnt5A (B) and NF-κB (C) were examined by real time PCR. GAPDH is internal control for equal calculation. *P < 0.05