FIGURE 1.

LncRNA HHAS1 is upregulated during the osteogenic differentiation of BMSCs. (A) The heat map of microarray showed the differently expressed lncRNAs of BMSCs 10 days after osteogenic induction. (B) The relative expression levels of lncRNA HHAS1 were upregulated during the osteogenic differentiation of BMSCs and positively correlated with the mRNA expression of OCN, Osterix, ColI, ALP and the intensity of ARS staining. (C) Schematic annotation of the lncRNA HHAS1 genomic locus on chromosome 4 and the conservation analysis of HHAS1 by Phylop on the UCSC browser. The rectangles represent exons. (D) The coding potential was analyzed by CPC and CPAT, which indicated that lncRNA HHAS1 had very low coding potential. (E) The intracellular localization of lncRNA HHAS1 in BMSCs was measured by RNA‐FISH assays. U6 was the nuclear control, and 18S was the cytoplasmic control (scale bar = 10 μm). (F) The percentages of nuclear and cytoplasmic RNA were measured by qPCR after subcellular BMSCs fractionation. U6 and MALAT1 served as nuclear controls, GAPDH served as the cytoplasmic control, and GAS5 served as both a nuclear and cytoplasmic control. The correlation data in (B) were determined by Pearson correlation and linear regression analysis (n = 15). Other results are presented as the mean ± SD (n = 10, determined by independent‐sample t‐tests). All experiments were performed three independent times, *p < 0.05, **p < 0.01