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Figure 6 The majority of Zrf1 positive cells are non-proliferative and do not incorporate BrdU. (A–F) anti-BrdU/anti-Zrf1 double immunofluorescence in transverse cryosection of 20 µm of the POA. Treatment with two pulses of BrdU at 1 and 7 days, and the brains were fixed at 8 days. Zrf1+ tanycytes (red), generally do not express BrdU (green), but a low number of Zrf1+ cells are BrdU+ ((B), magnified image of boxed). All sections were labeled with DAPI (blue). Scale bar: 30 μm. (G) Percentage of BrdU+, Zrf1+, and Zrf1+/BrdU+ cells distributed in the wall of the DiV of the POA. The cell count was obtained of the total immune-labeling cells of the POA. (H) Percentage of BrdU+, Zrf1+, and BrdU+/Zrf1+ cells are located in representative sections of the POA (PPa1 = 2, PPa2 = 2, PPa3 = 2, PPa4 = 3 and PPp1 = 2) where BrdU+, Zrf1+, and Zrf1+/BrdU+ cells were counted.