Distribution of EGFP in hypothalamic glial cells. (A–F) Representative confocal images of one focal plane using anti-vimentin (red) or anti-GFAP (red) antibodies as well as the fluorescence associated with EGFP (green) and the nuclear marker, TOPRO-3 (blue). (A–C) Adenoviral transduction in α-tanycytes. (D–F) Adenoviral transduction of β-tanycytes. (A,C) EGFP detected in apical membranes (arrows) and in long cellular processes in the basal hypothalamus (arrowheads). (B,E) Vimentin showing both proximal (arrows) and distal (arrowheads) tanycyte regions and colocalization with the EGFP. (C,F) GFAP showing astrocyte distribution, the absence of GFAP in the ventricular wall (arrows) and EGFP-positive processes (arrowheads). Scale bars (A–H) 50 µm.