Figure 1

Glucose deprivation and 2-DG treatment reduce miR-483-3p expression in HepG2 cell lines. (a) MiR-483-3p relative expression by RT–qPCR in HepG2 cells cultured in Dulbecco’s modified Eagle’s medium (DMEM) media without glucose (black bars) or with glucose 10 mM (gray bars) for 10, 20, 36 and 48 h. (b) Relative luciferase activity of the miR-483 promoter sequence containing the E-Box interacting with CTNNB1/USF1 complex, in HepG2 cells cultured in DMEM media without glucose (black circles) or with glucose 10 mM (gray circles) for 0, 16 and 36 h. (c) MiR-483-3p, IGF2 (left y axis) and primiR-483 (right y axis) relative expression by RT–qPCR in HepG2 cells treated with 2-DG at 2 and 10 mM for 48 h. MiR-483-3p expression was normalized on U44, whereas IGF2 and primiR-483 on ACTB. (d) Relative luciferase activity of the miR-483 promoter sequence containing the E-Box interacting with CTNNB1/USF1 complex, in HepG2 cells treated with 2-DG 5 mM in low (black bars) or high (white bars) glucose condition (1 and 4.5 g/l, respectively) for 48 h. As control for the wild-type vector (wt) was used mutated vector (mut) for the region interacting with the CTNNB1/USF1 complex. The graphs represent the means of technical triplicates with the respective s.d. For statistical analysis, Student’s t-test was applied (*P<0.05; **P<0.01; ***P<0.001; ****P<0.0001).