Research Paper Volume 11, Issue 1 pp 18—32

Figure 5. ILK is a target of miR-542-3p. (A) Volcano plot showed the variation in gene expression. The negative log of adj.P.Val (base 10) is plotted on the y-axis, and the log of the FC (base 2) is plotted on the x-axis; (B) Heat map of differentially expressed mRNAs in normal and osteosarcoma tissues; (C) ILK expression in osteosarcoma tissues were examined by qRT-PCR. *P<0.05 compared to the peri-tumorous group; (D) Scatter diagram exhibited a negative correlation of ILK and miR-542-3p in 20 pairs of osteosarcoma tissues by qRT-PCR; (E) The binding site in miR-542-3p and 3'-UTR of ILK were indicated by TargetScan; (F) Luciferase reporter assay data found that co-transfection of osteosarcoma cells with miR-542-3p mimics and wild-type (WT) ILK 3'-UTR significantly decrease the luciferase activity, whereas co-transfection with mutant-type (MUT) ILK 3'-UTR and miR-542-3p mimics showed no difference with the control group; (G) Western blot was used to tested the expression of ILK in the normal human osteoblastic cell line hFOB1.19 and the human osteosarcoma cell lines 143B and U-2OS; (H) RT-PCR was used to quantify the endogenous levels of ILK in hFOB1.19, 143B and U-2OS. **P<0.01 compared with the hFOB1.19 group; (I) RT-PCR was used to measure ILK expression levels after transfection of a pcDNA3.1-ILK and si-ILK in 143B and U-2OS cells. *P<0.05, **P<0.01 compared with the NC group. (J) ILK expression was inhibited by miR-542-3p mimics (mimics group), which reversed by miR-542-3p inhibitor (inhibitor group). **P<0.01 compared with the NC group.