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Fig. 4 | Stem Cell Research & Therapy

Fig. 4

From: Long non-coding RNA SNHG5 promotes the osteogenic differentiation of bone marrow mesenchymal stem cells via the miR-212-3p/GDF5/SMAD pathway

Fig. 4

SNHG5 serves as a sponge for miR-212-3p. a Confocal FISH images show the subcellular localization of SNHG5 in hBMSCs (n = 3). Nuclei were stained by DAPI (blue). Scale bars: 20 μm. b Venn diagrams show the number of potential miRNAs targeting SNHG5 (including miR-212-3p). The potential miRNAs were predicted by three databases: LncBase Predicted v.2, starBase, starBase v2.0. p. c Volcano plot shows that a total of 56 genes were significantly upregulated and 27 genes were significantly downregulated in SNHG5 knockdown hBMSCs. The DEGs were defined as fold change ≥ 2 and p < 0.05. d Percentage of nuclear and cytoplasmic RNA levels of SNHG5 and miR-212-3p (n = 3). MALAT1 and U6 were used as fractionation indicators. e, g Relative RNA expression of miR-212-3p after downregulation or upregulation of SNHG5 or YY1 by qRT-PCR (n = 3). U6 was used for normalization. f Schematic diagram of SNHG5 WT and SNHG5 Mut luciferase reporter plasmids. A putative miR-212-3p target site of SNGH5 was predicted in bioinformatics analyses. h, i Dual luciferase reporter assays validate the interaction between miR-212-3p and SNHG5 in 293 T cells and hBMSCs (n = 3). (*p < 0.05; **p < 0.01; ***p < 0.001). DEGs, differentially expressed genes; FISH, fluorescence in situ hybridization; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; hBMSC, human bone marrow mesenchymal stem cells; MALAT1, metastasis-associated lung adenocarcinoma transcript 1; qRT-PCR, quantitative reverse transcription-polymerase chain reaction; SNHG5, small nucleolar RNA host gene 5

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