(A) Cartoon schematic outlining prospective isolation from dissection to cell identification. (B, C) Biaxial gating strategy to obtain V-SVZ cell types. Symbols at terminal gates correspond to cell types in (C). To the right are representative histograms showing intensity of p-4EBP1 T37/46 signal in freshly isolated dorsal (blue) and ventral (red) TAPs. Black histogram outline shows the fluorescence minus one control for each sample. (C) Graphs show median fluorescence intensity data for p-4EBP1 T37/46 from 7 sets of 15 pooled mice each (105 total mice); left: standard scale and right: arcsinh-transformed. p-4EBP1 T37/46 is elevated in ventral TAPs relative to dorsal (P = 0.0175, paired t test, bar = median) but is not different in the other cell types shown (qNSCs [P = 0.1931], aNSCs [P = 0.5853], and neuroblasts [P = 0.1374]). (D) Representative tilescan confocal images of V-SVZ stained for DAPI (blue), Mash1 (red), and p-4EBP1 T37/46 (green), with boxed areas highlighted to the right. Scale bars: 100 μm (tilescan), 10 μm (63× images). (E) Quantification of per-cell intensity for p-4EBP1 T37/46 (top: 1,026 dorsal and 590 ventral cells total) and p-S6 S240/244 (bottom: 1,306 dorsal and 805 ventral cells) in confocal images. Both phosphorylation events are elevated in ventral Mash1+ cells relative to dorsal (P < 0.0001 [p-4EBP1] and P = 0.0161 [p-S6 S240/244], Wilcoxon signed rank tests). Each experiment: n = 4 mice, 3 sections/mouse. For all graphs, bars represent mean ± SD.