Figure 3

Pax6^-/- mutant ES cells generate embryoid bodies containing normal proportions of proliferating and differentiating cells. (A) Histograms show percentages of cells positive for either BrdU, β-III-tubulin or both two hours after plating cells from dissociated EBs that had been exposed to a short pulse of BrdU shortly before dissociation. From left to right: percentages of all cells that were BrdU positive on Day 6 after LIF withdrawal; percentages of all cells that were BrdU positive on Day 8; percentages of all cells that were β-III-tubulin positive on Day 8; percentages of BrdU positive cells that were β-III-tubulin positive on Day 8. All values are means of 3 or 4 independent culture runs ± standard errors. No significant difference was observed between the wild type and Pax6^-/- mutant ES cell lines within each culture paradigm. (B) Examples of wtMM4 cells that are BrdU positive (red), β-III-tubulin positive (green), double-labeled (yellow arrow) and labeled only with TOPRO3 counterstain (blue).