Figure 1
From: MiR-29b controls fetal mouse neurogenesis by regulating ICAT-mediated Wnt/β-catenin signaling
MiR-29b reduces NSC proliferation. (a and b) NSCs were harvested and the expression of miR-29 subtypes a, b, and c was quantified by qRT-PCR. MiR-29b, but not miR-29a or c, showed a prominent (approximately sixfold) increase following induction of differentiation. ICAT was highly expressed in fetal NSCs, but its expression declined with differentiation. (c and d) Fetal NSCs transfected with miR-29b or anti-miR-29b were plated on laminin-coated dishes, cultured in medium supplemented with EGF and FGF2 for 24 h, and labeled with a 1-h pulse of BrdU. Cells exhibiting a BrdU signal in DAPI-stained nuclei were considered positive. Approximately 1000 cells were quantified for each section. (e) Fetal NSCs transfected with miR-29b or siICAT were analyzed by qRT-PCR. Nestin expression was reduced by ~90% in ICAT-deficient NSCs. (f) Fetal NSCs transfected with miR-29b were analyzed by qRT-PCR and western blotting (1, siCONTROL; 2, miR-29b). The miR-29b transfection remarkably inhibited ICAT protein expression but ICAT mRNA was reduced by only ∼15% in NSCs (P=0.0525, Student’s t-test, f). Data are expressed as mean±S.E.M. (error bars) of triplicate experiments (*P<0.05, **P<0.01, ***P<0.005; Student’s t-test)
