Wide cell transduction is achieved in the brain stem and cerebellum of nonhuman primates that were kept in the Trendelenburg position for 5 or 10 minutes. (a–f) DAB staining showed that several areas of the brainstem were transduced by scAAV9.CBA.GFP compared to the negative control (a–c). In particular, the XII and the V cranial nerve nuclei, the hypoglossal (b–e) and the trigeminal (c–f) nucleus respectively, were highly targeted. (d) Upper motor neuron counts revealed that about 50% motor neurons were transduced in the V layer of the motor cortex, and more than 60 and 75% motor neurons were GFP+ in the hypoglossal and trigeminal nucleus respectively; GFP (green) and ChAT (red). (g–i) High transduction levels were also achieved in the cerebellum, in particular the Purkinje cells and the nuclear layer. Error bars = SD; n = 5.