Isolation, profiling, and QISP analysis of GFP + (Eomes+) cortical neurons. a, Neocortex (NCX) containing GFP + neurons is dissected, and cells isolated with flourescence activated cell sorting (FACS). Following transcriptional profiling, genes are sorted by RMA and their level of fold up- or down-regulation compared to the FACS GFP- population. Verification of up-regulated genes is accomplished with quantified in situ hybridization (QISP), and the verified genes hierarchically clustered (see Methods). b, Fluorescence microscopic images of a section from an E14.5 Eomes::eGFP brain, displaying GFP + (green) and GFP- cells, that are blue due to Hoechst nuclear labeling. Subregions of Interest (sROIs) 1-20 are superimposed in white on the image. Scale bar, 25 μm. MZ, marginal zone; CP, cortical plate; IZ, intermediate zone; SVZ, subventricular zone; VZ, ventricular zone; LGE, lateral ganglionic eminence. c, Representative outcome of FAC sort of GFP + and GFP- cells. A GFP + (P3) and GFP- (P4) were isolated based on fluorescence (GFP-A) and side scatter (SSC-A). The boxes denote the relevant sorted populations. All in situ hybridizations were obtained from Genepaint.
Cameron et al. BMC Neuroscience 2012 13:90 doi:10.1186/1471-2202-13-90