Fig. 3: Inhibition of miR-652-5p on Tigar expression in T-ALL cells.
From: MiR-652-5p elevated glycolysis level by targeting TIGAR in T-cell acute lymphoblastic leukemia
(A) Conserved binding site for miR-652-5p within the Tigar mRNA 3ʹUTR as predicted by TargetScan software. Top, structural features of Tigar. The predicted binding site (bs) within the Tigar 3ʹUTR for miR-652-5p is indicated by inverted triangle. Middle, the sequence of miR-652-5p. Solid line indicated the first binding region, dotted line indicated the second binding region. Bottom, the sequence of fragment which is insert into plasmid for luciferase reporter assay. Relative luciferase activity in 293 T cells cotransfected with miR-652-5p and reporter or control luciferase plasmids containing the wildtype sequence of Tigar 3ʹUTR (B) or containing the mutated sequence (C). (D) Relative luciferase activity in 293 T cells cotransfected with miR-652-5p mimics and different reporter plasmid. The expression of Tigar mRNA (E) and protein (F) in Jurkat and Molt-4 cells with impaired miR-652-5p. Top indicated the representative image of Western Blot. Bottom indicated the statistics and normalized to control as 1.0 in F. Pos1., the binding position 1 for miR-652-5p to Tigar mRNA; Pos2., the binding position 2 for miR-652-5p to Tigar mRNA WT, the reporter plasmid with wildtype sequence of Tigar mRNA; MUT, the reporter plasmid with mutated sequence of Tigar mRNA; B blank; V, MiR vector with miR-652-5p; U1, vector with the first binding sequence for miR-652-5p to Tigar mRNA; U2, vector with the second binding sequence for miR-652-5p to Tigar mRNA; MU1, the mutated U1; MU2, the mutated U2; Scram, control; Si, impaired miR-652-5p; J, Jurkat cell lines as control; M, Molt-4 urkat cell lines. p is compared to control and calculated by t student; *p < 0.05; **p < 0.01. **/** indicated that the group was compared to V + MIR group/to V + U1or U2 group.
