Fig. 4

TUG1 targeted on miR-34b-5p in the models of sepsis. (a) The putative binding site between TUG1 and miR-34b-5p were predicted on miRDB. The wild-type binding sequence and the designed mutant sequence were indicated as TUG1-WT and TUG1-MUT, respectively. (b) PMVECs were transfected with miR mimics or miR-34b-5p mimics. The transfection efficacy was determined by measuring the level of miR-34b-5p in transfected cells. (c) HEK-293 T cells were co-transfected with miR mimics (or miR-34b-5p mimics) and TUG1-WT (or TUG1-MUT). The luciferase activity was measuring 48 h post transfection using dual-luciferase reporter assay. (d) The RIP assay was performed in PMVECs using a mouse monoclonal anti-Ago2 antibody as a positive control and an anti-IgG antibody as a negative control. The expressions of TUG1 and miR-34b-5p in RISCs were analyzed using qRT-PCR. (e) The relative expression of miR-34b-5p in PMVECs transfected with control adenoviral vector (Vector) or adenoviral vector expressing TUG1(TUG1) were examined using qRT-PCR. (f) The relative expression of miR-34b-5p in sham- and CLP-operated mice were detected using qRT-PCR. (g) The correlation between miR-34b-5p expression and TUG1 level in mouse lung tissues were analyzed using Spearman’s rank correlation test