Figure 1
Expression pattern of miR-214 in the developing neocortex and cultured neural stem cells. (A) Predicted secondary structure and sequence conservation of the miR-214 precursor hairpin. The maturation regions of miR-214 and miR-214* are marked with red straight lines, and the colour of the base represents its conservation with higher conservation in red and lower in blue (modified from Rfam database 11.0). (B) Microarray analysis of the expression level of the mature miR-214 and miR-214* in the mouse embryonic cerebral cortex. Total RNAs of the E12.5, E14.5, E16.5 and E18.5 dorsal cortexes were extracted and tested by the LC Sciences company. Error bars show the standard error of mean. (C) The expression dynamics of miR-214 and miR-214* in the mouse embryonic cerebral cortex. In situ hybridization for miR-214 and miR-214* on coronal sections of the embryonic telencephalon. The top left panel of each stage shows the expression of miR-214, and the top right panel shows that of miR-214*. The bottom panel shows the corresponding neocortical areas of the black dashed boxes of the top left panel at a higher magnification. VZ, ventricular zone; SVZ, sub-ventricular zone; IZ, intermediate zone; CP, cortical plate; the scale bar in C is 100 μm. (D,E) Immunofluorescence of NSCs during proliferation conditions (D) and the differentiation stage (E). (D) Staining of the neural sphere with neural stem cell markers Pax6 (green) and Nestin (red) (E) Immunofluorescence of differentiated cells 3 days after induction with neuron marker MAP2 (Red) and astrocyte marker GFAP (green). Scale bar: 100 μm. (F) Expression levels of miR-214 and miR-124 before and after differentiation of NSCs. Total RNA of the NSCs and differentiated cells was extracted and tested by real-time PCR, and the miR-214 signal was normalized to that of the U6 snRNA (n = 3, p = 0.035).
