Fig. 3: miR-184-3p directly modulates the expression of CRTC1.

a Top panel: graphical representation of CRTC1 mRNA sequence (ENST00000338797.6) showing 3’UTR and predicted hsa-miR-184-3p binding sites (in red); bottom panel: nucleotide position of hsa-miR-184-3p binding sites and sequence context within the 3’UTR region of CRTC1 mRNA. b Luciferase assay in HeLa cells transfected with a plasmid overexpressing hsa-miR-184-3p compared to cells transfected with a scrambled vector; values are given as mean ± SD of relative luciferase activity compared to control, after normalisation of firefly luciferase unit to renilla luciferase activity; statistics using Mann–Whitney U test (n = 4 independent experiments). c RT-Real-time PCR analysis of CRTC1 expression in EndoC-βH1 cells transfected with a synthetic inhibitor of hsa-miR-184-3p; data are expressed as fold change compared to scrambled control; Mann–Whitney U test statistics (n = 6 independent experiments). d Western blot analysis of CRTC1 (78 kDa) in EndoC-βH1 cells transfected with a synthetic inhibitor of hsa-miR-184-3p. Data are presented as mean ± SD of fold-change values of normalised CRTC1/β-Act ratio of miR-184-p inhibitor vs. miR-Ctr inhibitor-transfected samples; statistics using paired Student’s t-test (n = 6 independent experiments).