Figure 2
From: miR-200 family controls late steps of postnatal forebrain neurogenesis via Zeb2 inhibition
MicroRNA expression in OB subpopulations.
(a) Sagittal section of a GAD67-GFP knock-in mouse forebrain. Diagrams are dot plots of the FACsorting experiment performed on wild-type (left) and GAD67-GFP knock-in mice (right). Three cell populations were sorted from GAD67-GFP knock-in mouse brain and used for subsequent qRT-PCR analyses. (b) qRT-PCR characterization of the three populations. GluR2 is expressed on both, GABAergic neuronal progenitors and fully differentiated neurons. Doublecortin is exclusively expressed in neuronal progenitors. GFP negative cells do not significantly express these neuronal markers. (c) qRT-PCR analysis of the expression of miR-200b and miR-141 in the three sorted populations showing a preferential expression in the GFP-low fraction. (d) qRT-PCR characterization of the two purified cell fractions issued from the MACS experiment designed to discriminate neuronal vs glial fraction from the OB based. Neuronal (NeuN, GluR2, DCX) and glial (GFAP, Olig1, Olig2) markers validate the expected neuronal and glial identities. (e) qRT-PCR analysis demonstrates that miR-200b and miR-141 are enriched in the neuronal fraction. For b-e the qPCR values shown in the histograms result from 2 (b,d) or 3 (c,e) qPCR experiments (4 wells per condition in each experiment) (f) Electroporation of an expression construct driving GFP with regulatory sequences of the human miR-429/miR-200a/miR-200b cluster leads to GFP-labeled cells in the OB. Scale bar: 70 μm. Error bars: sem.
