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. 2018 Oct 3;24(4):588–600. doi: 10.1038/s41380-018-0243-x

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — Laser capture microdissection (LCM) and RNA-sequencing of pyramidal neurons and non-pyramidal cells. a Representative micrographs highlighting Nissl-stained frozen tissue (top) and NeuN immunohistochemistry (bottom). Arrows indicate pyramidal neurons and arrowheads indicate adjacent non-pyramidal, NeuN- cells. b Scatter plot displaying genes expressed and statistically enriched in pyramidal (purple) and non-pyramidal (orange) populations (P < 0.01). c Scatter plot displaying relationship between pyramidal enrichment pattern (x axis, LCM) and neuronal enrichment (y axis, sorted mouse neocortex [44]). Thin line indicates regression with all genes and thick line indicates regression of with only differentially expressed (colored) genes. Counts in each corner indicate number of genes within the respective quadrant that were both statistically enriched (P < 0.01 based on LCM) and showed at least a 2²-fold enrichment pattern (based on sorting). d Data from Zhang et al. [44]. highlighting the enrichment pattern of Neurod6 and Lrp4, enriched in neuronal and non-neuronal cells, respectively. e In silico cytometry revealed enrichment of neuronal cells within the pyramidal population and a relative depletion of this cell type in the non-pyramidal population (P < 0.05). f Non-linear cluster analysis highlighted the topological relatedness between the pyramidal population (purple) and excitatory cortical neurons (blue) from the human neocortex, especially the Ex1 subtype (dark blue), which reflects layers II and III cortical projection neurons as defined by Lake et al. [45]. The non-pyramidal cells (orange), on the other hand, were more similar to glia (green) and inhibitory cortical neurons (red)