Fig. 2
MiR-153 targets the 3’UTRs of the Snap25, Vamp2, Stx1a and Syt1 genes. a The levels of miR-153, miR-137, miR-34c and miR-135a in the hippocampi of sham and 2VO rats as assessed by qRT-PCR (for miR-153: PLevene = 0.008, Student’s t-test: P = 0.011), n = 6. bMiR-153 levels in the plasma of VaD patients as assessed by qRT-PCR (PLevene = 0.002, Student’s t-test, P = 0.019). VaD: vascular dementia. c Complementarity between the miR-153 seed sequence and the 3’UTRs of Snap25, Vamp2, Stx1a and Syt1 predicted using the TargetScan5.1 algorithm and RNAhybrid database. The gene mutations are underlined. 3’UTR, 3′ untranslated region. d-g Dual luciferase assay of interactions between miR-153 and predicted binding sites or mutated binding sites in the 3’UTR of Snap25 (wild-type: PLevene = 0.192, one-way ANOVA: F = 85.093, P < 0.0001; 3’UTR mutant (3’UTR mut): PLevene = 0.184, one-way ANOVA: F = 0.199, P = 0.933), Vamp2 (wild-type: PLevene = 0.168, one-way ANOVA: F = 55.704, P < 0.0001; 3’UTR mut: PLevene = 0.041, one-way ANOVA: F = 2.561, P = 0.104), Stx1a (wild-type: PLevene = 0.276, one-way ANOVA: F = 119.138, P < 0.0001; 3’UTR mut: PLevene = 0.0331, one-way ANOVA: F = 0.155, P = 0.957) and Syt1 (Wild-type: PLevene = 0.522, one-way ANOVA: F = 825.5700, P < 0.0001; 3’UTR mut: PLevene = 0.077, one-way ANOVA: F = 1.603, P = 0.248) in HEK293T cells. n = 3 batches per group. Fisher’s PLSD test was used for the post hoc analyses of the two-group comparisons. *P < 0.05 vs control or psiCHECK™-2 vector; #P < 0.05 vs miR-153