Fig. 2
From: LncRNA CTD-2555A7.2 promotes bone formation with LncRNA-specific cascade amplification strategy
CTD-2555A7.2 bind with multiple miR-381-3p and promoted bone formation. A Sequence structure of CTD-2555A7.2.B miR-381-3p expression levels of hMSCs treated with CTD-2555A7.2 over-expression plasmid (compared with pCDNA3.1 plasmid), as detected by RT-PCR (mean ± SD, n = 3). vec: blank plasmid. CTD: plasmid containing CTD-2555A7.2 fell length sequence.C miR-381-3p expression levels of hMSCs treated with CTD-2555A7.2 siRNA (compared with negative control siRNA, mean ± SD, n = 3).D miR-381-3p expression levels of hMSCs treated with CTD-2555A7.2 full length, CTD-2555A7.2 non-binding region, CTD-2555A7.2 binding region and CTD-2555A7.2 mutant binding region over-expression plasmids respectively (compared with pCDNA3.1 plasmid), as detected by RT-PCR (mean ± SD, n = 3). non: plasmid containing CTD-2555A7.2 non-binding region. bind: plasmid containing CTD-2555A7.2 binding region. bind mut: plasmid containing CTD-2555A7.2 mutant binding region.E Binding effect of miR-381-3p and CTD-2555A7.2 binding region, as detected by luciferase reporter assay (mean ± SD, n = 3). Luc-mut: pMIR-Report luciferase reporter plasmid containing one mutant miR-381-3p binding repeat sequence of CTD-2555A7.2. Luc-WT: luciferase reporter plasmid containing one wild-type miR-381-3p binding repeat sequence of CTD-2555A7.2. Ago-NC: cells treated with agomiR-NC. Ago-381-3p: cells treated with agomiR-381-3p.F,G Binding effect of miR-381-3p and CTD-2555A7.2 different regions (compared with pMIR-Report luciferase reporter plasmid) and treated by agomiR-381-3p (left) or antagomiR-381-3p (right), as detected by luciferase reporter assay (mean ± SD, n = 3). Atgo-NC: cells treated with antagomiR-NC. Atgo-381-3p: cells treated with antagomiR-381-3p.H ALP and Alizarin Red staining of hMSC cell treated with CTD-2555A7.2 exprssion plasmids of different regions, as detected by ALP staining and Alizarin Red staining. I. TCF7 activities of hMSCs treated with CTD-2555A7.2 exprssion plasmids of different regions, as detected by luciferase reporter assay (mean ± S.D., n = 8).J,K Calvarial bone mineral apposition rates of C57BL/6 mice treated with CTD-2555A7.2 exprssion plasmids of different regions (mean ± SD, n = 4). Scale bar: 10μm.*P < 0.05, **P < 0.01, ***P < 0.001
