Research Paper Volume 16, Issue 4 pp 3896—3914

GAS5, a long noncoding RNA, contributes to annulus fibroblast osteogenic differentiation and apoptosis in intervertebral disk degeneration via the miR-221-3p/SOX11 axis

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Figure 5. The role of GAS5 in the apoptosis and osteogenic differentiation of AFs was mediated by SOX11. (A) AFs were transfected with SOX11 overexpression or an empty vector, and the transfection efficacy was detected using qRT-PCR (n = 3). Next, AFs were transfected with or without si-GAS5 plasmid alone or with the SOX11 overexpression vector. (B) ALP and (C) ARS staining assays were used to evaluate the osteogenic differentiation level (n = 3). (D) The osteogenic differentiation-related markers were detected using western blotting (n = 3). (E) The apoptosis-related protein levels were measured using western blotting (n = 3). (F) TUNEL staining assay was employed to evaluate the apoptosis of AFs (n = 3). (*p < 0.05, **p < 0.01, and ***p < 0.001, vs. the vector group; #p < 0.05, ##p < 0.01, and ###p < 0.001, vs. the si-GAS5 group).