Depleted Long Noncoding RNA GAS5 Relieves Intervertebral Disc Degeneration via microRNA-17-3p/Ang-2

Figure 5

GAS5 upregulates Ang-2 expression in NPCs through downregulating miR-17-3p expression. (a) The binding sites between Ang-2 and miR-17-3p predicted using RNA22 website (https://cm.jefferson.edu/rna22/Interactive/). (b) The luciferase activity of Ang-2 reporter plasmids determined by dual-luciferase reporter assay ( vs. HEK 293T cells treated with NC mimic). (c) Expression of Ang-2 in NPCs after miR-17-3p was upregulated or downregulated and determined by RT-qPCR, which was normalized to GAPDH. (d, e) Western blot analysis of Ang-2 protein in NPCs after miR-17-3p was upregulated or downregulated, which was normalized to GAPDH. (f) Correlation analysis of miR-17-3p expression and Ang-2 expression in NP tissues. (g) Expression of Ang-2 in NPCs after GAS5 was upregulated or downregulated and detected by RT-qPCR, which was normalized to GAPDH. (h, i) Western blot analysis of Ang-2 protein in NPCs after GAS5 was upregulated or downregulated, which was normalized to GAPDH. vs. HDNPCs treated with si-NC, ^# vs. HDNPCs treated with oe-NC, and ^$ vs. HDNPCs treated with oe-GAS5. Data () were analyzed by one-way ANOVA, followed by Tukey’s post hoc test, while data in (b) were analyzed by unpaired test. The cell experiment was repeated independently 3 times.

(a)

(b)

(c)

(d)

(e)

(f)

(g)

(h)

(i)