Nature Protocols: doi: /nprot Supplementary Figure 1. Effect of different enzymes on CD13 and CD31 populations.

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Gervais Whitehead
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Supplementary Figure 1 Effect of different enzymes on CD13 and CD31 populations. Comparison of cortex dissociated for 30 minutes with collagenase/dispase digestion (a) or papain (b).

1 Supplementary Figure 1 Effect of different enzymes on CD13 and CD31 populations. Comparison of cortex dissociated for 30 minutes with collagenase/dispase digestion (a) or papain (b). Papain degrades the CD31 epitope and CD13 + pericytes are greatly reduced (b). Percentages refer to the proportion of cells in the previous parent gate. Plots show 50,000 events. Experiments were performed with the approval of Columbia University IACUC and the cantonal veterinary office Basel-Stadt.

2 Supplementary Figure 2 FACS controls for setting gating strategy. Representative plots for isotype, single colour and FMO controls for FACS strategy. Isotype controls and the percentage of non-specific labeling for each fluorophore are shown in a (FITC), b (PE), and c (APC). Unstained cells (d-e), single colour controls: CD45-PE and CD41-PE (f); CD31-APC (g); CD13-FITC (h) and FMO controls (CD13-FITC (i); CD31-APC (j)) are used to set gating strategy. Plots show 50,000 events. Experiments were performed with the approval of Columbia University IACUC and the cantonal veterinary office Basel-Stadt. a c adapted with permission from Crouch et al. (ref. 21), Regional and stage-specific effects of prospectively purified vascular cells on the adult V-SVZ neural stem cell lineage, J. Neurosci., vol. 35, Copyright 2015; permission conveyed through Copyright Clearance Center.

3 Supplementary Figure 3 Schema of dissection of cortex. Overview of dissection of cortex from the brain. Steps refer to numbers in Box 1. First, remove the meninges from brain surface. Make four coronal cuts in the brain. Dissect the cortex from each coronal section. Use the corpus callosum (black) as a ventral limit to dissect the cortex (dashed white line).

4 Supplementary Figure 4 Dissection of the mouse cortex. (a-b) Dorsal view of whole mouse brain before (a) and after (b) removal of the meninges. (c-d) Coronal slice of the mouse brain before (c) and after (d) dissecting the cortex. Experiments were performed with the approval of Columbia University IACUC and the cantonal veterinary office Basel-Stadt.

5 Supplementary Figure 5 Schema of dissection of V-SVZ tissue. Overview of V-SVZ dissection. Steps refer to numbers in Box 2. To dissect the V-SVZ from the brain, make 4 coronal cuts through the brain and discard the most rostral and caudal sections. From each coronal section, dissect the V-SVZ (thin brown wedge beside the lateral ventricle) by cutting ventrally below the ventricle (step 3), dorsally by pulling up on the corpus callosum (step 4), and laterally to remove striatum (step 5). Repeat on the contralateral side before proceeding to the next slice.

Supplementary Figure 6 Dissection of the mouse V-SVZ. (a) Photograph of coronal section of mouse brain showing bilateral V-SVZ outlined by dashed lines.

6 Supplementary Figure 6 Dissection of the mouse V-SVZ. (a) Photograph of coronal section of mouse brain showing bilateral V-SVZ outlined by dashed lines. (b-c) The V-SVZ has a distinct, more velvety texture than the adjacent striatum, which contains white matter tracts. The V-SVZ is located to the right of the dashed line in (c) adjacent to the lateral ventricle. Cc, corpus callosum; str, striatum. Experiments were performed with the approval of Columbia University IACUC and the cantonal veterinary office Basel-Stadt.

Chapter 3. Structure and Function of the Nervous System. Copyright (c) Allyn and Bacon 2004

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