Figure 4

Pax6^-/- ES cells generate abnormally low proportions of neurons. (A-F) Examples of cultures of cells derived from wtMM4, SeyD1 and SeyD2 lines on Day 14 after LIF withdrawal stained with either β-III-tubulin or MAP2 (green) and TOPRO-3 (blue). (G) Mean percentages (± standard error of the mean; n = 3-4 independent cultures per line) of β-III-tubulin (open bars) or MAP2 (grey bars) expressing neurons are shown for each ES cell line. Percentages of β-III-tubulin-positive and MAP2-positive neurons were significantly reduced in Pax6 mutant ES cell lines SeyD1 and SeyD2 (with Student's t-tests, differences for β-III-tubulin were both p < 0.0001 and for MAP2 were both p < 0.02).