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. 2017 Aug 30;51(4):1115–1123. doi: 10.3892/ijo.2017.4110

Figure 2.

Figure 2

miR-144-3p is regulated by TUG1 in osteosarcoma cells. (A) The expression of miR-144-3p was analyzed in osteosarcoma tissues compared with adjacent normal tissues by RT-qPCR (n=25, P<0.0001). (B) miR-144-3p expression levels were determined by RT-qPCR in osteosarcoma cell lines. (C) The correlation between TUG1 and miR-144-3p expression was determined in osteosarcoma tissues. (D) miR-144-3p was upregulated when TUG1 was knocked down. (E) Sequence alignment of miR-144-3p and TUG1 (in red). miR-144-3p reduced the activity of the luciferase reporter with the specific sequence of TUG1 but not with the mutant sequence. (F) Amount of TUG1 bound to Ago2 or IgG measured by RT-qPCR after RIP, IgG was used as a negative control and SNRNP70 was used as a positive control. (G) U2OS cells were transfected with biotinylated WT miR-144-3p (miR-144-3p-Bio) or biotinylated mutant miR-144-3p (miR-144-3p-Mut-Bio) or biotinylated NC (NC-Bio). Forty-eight hours after transfection, cells were collected for biotin-based pull-down assay. TUG1 expression levels were analyzed by RT-qPCR. Error bars indicate s.d. *P<0.05, **P<0.01, Student's t-test.