Figure 2.

rAAV4 Vectors with Distinct Promoters Reveal Differential Selectivity for rNSCs

(A) Confocal imaging of AAV4-CMV-mCherry-transduced cells stained for RFP, GFAP, and DCX. z-Stack confocal data were maximum-intensity projected in ImageJ.

(B) 3D reconstruction of a deconvoluted confocal image in (A) using Huygens Professional software.

(C) Six cells were classified for cell identity based on morphology and cell stage markers.

(D) Confocal images showing marker expression of cells (from A) at a single focal plane. Three RFP⁺ cells (cells 1, 3, 5) with radial processes expressed GFAP but not DCX. These cells were classified as rNSCs. One non-radial precursor cell (cell 6) expressed GFAP but not DCX. Cells of this stage were classified as early progenitor cells (EPCs). Two RFP⁺ cells (cells 2 and 4) did not express GFAP and DCX. These cells were classified as intermediate progenitor cells (IPCs).

(E and F) Confocal images showing the DCX expression in RFP⁺ cells transduced by rAAV4-CBA-tdTomato (E) and rAAV4-CMV-mCherry (F). Note some rAAV4-CBA-tdTomato-transduced RFP⁺ cells were DCX⁺ (arrowhead), and some were DCX⁻ (asterisks); in contrast, most rAAV4-CMV-mCherry-transduced RFP⁺ cells were DCX⁻.

(G) Quantification of the cell types found in CBA-injected and CMV-injected mice at 24 hpi (n = 3 mice).

(H–J) rAAV4-CMV-mCherry displayed similar tropism for rNSCs in CD-1 mice compared with C57BL/6 mice at 3 days post injection (dpi) (n = 3 mice).

Scale bars, 20 μm. Values represent mean ± SD. See also Figures S2 and S3.