Lineage Is a Poor Predictor of Interneuron Positioning within the Forebrain  Christian Mayer, Rachel C. Bandler, Gord Fishell  Neuron  Volume 92, Issue 1, Pages 45-51 (October 2016) DOI: 10.1016/j.neuron.2016.09.035 Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 1 The Majority of Cortical Interneuron Clones in Mayer et al. Do Not Reside in Local Radial or Laminar Clusters (A) Schematic based on Figure S14 in Brown et al. (2011), illustrating the clonal organization of neocortical interneurons. Brown et al. concluded that cortical interneuron clones form spatially organized vertical and horizontal clusters. (B) From Figure S2 in Mayer et al. (2015). Reconstructions of cortical clones: front, side, and top view of 3D reconstructions illustrating the distribution of 16 cortical clones in the forebrain. The average distance (AD) of each clone is indicated. Neuron 2016 92, 45-51DOI: (10.1016/j.neuron.2016.09.035) Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 2 Density and 3D Distribution of Retrovirally Infected Interneurons in Mayer et al. and Brown et al. (A and B) Two experimental datasets from Brown et al. Three-dimensional reconstructions of the distribution of cortical interneurons in a postnatal Nkx2.1Cre/+;R26LSL-TVAiLacZ/+ mouse infected with retroviruses expressing EGFP. A dataset with 244 neurons that is representative of datasets used in Brown et al. 2011 (A) and a large dataset with 538 labeled interneurons (B) are shown. (C and D) Three-dimensional reconstructions of a representative dataset from Mayer et al. (2015) illustrating the distribution of cortical interneurons in a postnatal Nkx2.1Cre/+;R26LSL-TVAiLacZ/+ mouse that was infected with a retroviral library. The same dataset is shown before (C) and after (D) determination of clonal relations based on retroviral barcodes. Neuron 2016 92, 45-51DOI: (10.1016/j.neuron.2016.09.035) Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 3 The Majority of Cortical Interneurons Are Similarly Distributed Whether or Not They Share a Lineal Relationship (A) The nearest-neighbor distance (NND) decreases as the number of cells per dataset increases. In contrast, the average distance (AD) between pairs of neurons is not sensitive to the total number of data points per dataset. To illustrate this, NNDs and ADs were calculated for a high (200; blue dot) and a low (10; red five-pointed star) number of simulated data points (200; blue dot) in a given volume (the simulation was repeated 100 times). (B) Schematic illustration demonstrating how intra- and inter-clonal distances were calculated. The “intra-clonal distance” was calculated as the average distance between all possible pairs of clonally related interneurons (Bi). The “inter-clonal distance” was calculated as the average distance between all possible pairs of unrelated interneurons (Bii–iv), which includes the distance between (1) multi-cell clones (Bii), (2) single-cell clones (Biii), and (3) multi-cell clones and single-cell clones (Biv). (C) Box and whisker plot of the intra-clonal and inter-clonal distance. Whiskers indicate minimum to maximum values. All three datasets from Mayer et al. were included. The individual categories that made up all possible pairs of inter-clonal distance are shown. The number above the boxes indicates the n-number of interneuron pairs. (D) Scatter dot plot of the intra-clonal and inter-clonal distance broken down by brain (experimental dataset). No significance was observed when the intra-clonal and inter-clonal distances were compared (Kruskal-Wallis test, multiple comparison; Mann-Whitney nonparametric t test). Number above the boxes indicates the n-number of interneuron pairs. Data are presented as mean ± SEM. Relates to Figure 3F in Mayer et al. Neuron 2016 92, 45-51DOI: (10.1016/j.neuron.2016.09.035) Copyright © 2016 Elsevier Inc. Terms and Conditions