Supplementary Figure 8: Cre immunostaining establish GFP specificity of CRE-DOGOPT in rAAV-infected cerebella (Related to Figure 6).
From: Cell type–specific manipulation with GFP-dependent Cre recombinase
2/1 serotype rAAV-CRE-DOGOPT and rAAV-FLEX-tdT were injected into the cerebellum of Tg(GAD67-GFP)+ or GFP negative animals and harvested 3 weeks later. Same experiment as in Fig. 6. (a) Top panel: Images of rAAV-injected lobule V in Tg(GAD67-GFP)+ animal. Anti-Cre immunostain and GFP-dependent expression of tdT indicate successful virus injection. Bottom Panel: Images of uninfected lobule in the same animal show no expression of Cre and tdT. Scale bar, 50 μm (b) Top Panel: Images of rAAV injected lobule V in Tg(GAD67-GFP) negative animal. Anti-Cre immunostain indicates successful infection, but tdT is not expressed in the absence of GFP. Bottom Panel: Images of uninfected lobule in the same animal shows no expression of Cre and tdT. In all Merged panels, green is GFP, blue is Anti-Cre staining, red is tdT. Note the infected lobule panels in (b) were also shown in main text Fig. 6. All images are representative of 4 Tg(GAD-GFP)+ animals and 3 GFP-negative animals.
