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JSRM Code: 012010300005
SUPPLEMENTARY INFORMATION [PDF]

The Effect of Pro-Neurogenic Gene Expression on Adult Subventricular Zone Precursor Cell Recruitment and Fate Determination After Excitotoxic Brain Injury
Jones KS 1, Connor BJ1

Author Names in full: Kathryn S. Jones, 1, Bronwen J. Connor1

1Centre for Brain Research, Department of Pharmacology and Clinical Pharmacology, School of Medical Science, Faculty of Medical and Health Sciences, University of Auckland, 85 Park Road, Grafton, Auckland, New Zealand 1023.

Supplementary Figure


Supplementary Figure 1: Transcription factor expression in GFP-labelled cells in the anterior SVZ of RV-GFP control animals. (a) Image demonstrating the presence of OLIG2 (blue) expression in GFP-labelled (green) bipolar cells in the anterior SVZ (arrowheads) and in multipolar cells in the white matter and striatum (arrows). MASH1 was not observed in GFP cells in this image. (b) Image demonstrating the presence of DLX2 (blue) and MASH1 (red) expression in GFP-labelled bipolar neural precursor cells in the anterior SVZ (arrowheads). Scale bar a-b =50 µm

Supplementary Figure 2: Gene expression in GFP-labelled cells in the RMS of RV-GFP control animals. (a-c) Images demonstrating GFP-labelled cells within the central RMS are bipolar neuroblasts expressing DCX, that do not express MASH1 or OLIG2, but of which some are positive for DLX2. (a) GFP (green) / DCX (red). (b) MASH (red) / OLIG (blue) / GFP (green). (c) DLX (blue) / GFP (green). Arrowheads demonstrating the presence of GFP cells co-labelled with DLX2. Scale bar a-c =50 µm.

Supplementary Figure 3: DCX+ cells in the SVZ and Striatum do not label with NG2

Confocal z projection of (a-c) DCX (red) and NG2 (green). (d-f) represents boxed area in c at higher magnification. All DCX+ cells are negative for NG2 expression. Scale bar a-c = 200 µm and d-f = 50µm

SupplementaryFigure 4: Co-localisation of Pax6 transgene with RV-Pax6-GFP in the SVZ of control and QA lesioned animals. Confocal images of animals showing RV-Pax6-GFP cells (green) overexpressing PAX6 (red) in addition to endogenous NPCs expressing PAX6 in the SVZ of (a-c) unlesioned control animals (d-f) QA lesioned animals at 2 days post RV injection (dpi), (g-i) QA lesioned animals at 5dpi.  Arrows indicate some  of the co-labelled cells. Scale bar = 50 µm.

SupplementaryFigure 5: Co-localisation of Dlx2 transgene with RV-Dlx2-GFP in the SVZ of control and QA lesioned animals. Confocal images of animals showing RV-Dlx2-GFP cells (green) overexpressing DLX2 (red) in addition to endogenous NPCs expressing DLX2 in the SVZ of (a-c) unlesioned control animals (d-f) QA lesioned animals at 2 days post RV injection (dpi), (g-i) QA lesioned animals at 5dpi.  Arrows indicate co-labelled cells. Scale bar = 50 µm.

Supplementary Figure 6: Images of GFP-labelled cells in the RMS of non-lesioned RV overexpression animals. Images of GFP cells labelled in the RMS of control (no-lesion) animals after RV pro-neurogenic gene overexpression. Cells remain in the RMS and migrate to the olfactory bulb in (a) RV-Dlx2-GFP (scale = 100 µm) and (b) RV-Pax6-GFP (scale = 50µm) with no redirection into the striatum.

Supplementary Figure 7: GFP-labelled cells in the SVZ, RMS and striatum do not co-label with GFAP. (a-c) GFP labelled cells (green) in the SVZ/RMS of control animals are GFAP negative (red). (d-f) GFP labelled cells (green) recruited into the striatum of lesioned animals at are also GFAP negative (red). Scale bars = 25µm.