Figure 7: TSA treatment represses neural fate commitment of the anterior epiblast at E7.0.

(a) Immunocytochemistry for H3ac in E7.0 mouse embryos. Transverse sections from distal to proximal were shown. Scale bar, 75 μm. (b) Statistical analysis of H3ac⁺ cells in a (n=6). (c) Representative cellular morphologies of day 1 and day 5 cells differentiated in N2B27 medium from the anterior and posterior explants. The images were shown representatively for one of the three biological repeats. The arrows show the neurite outgrowth. Scale bar, 200 μm. (d) Gene expression analysis of the day 5 cells. Relative expression level was normalized by gapdh. (e) Representative cellular morphologies of the day 5 cells differentiated from the anterior explants with DMSO or TSA treatment. The images were shown representatively for one of three biological repeats. The arrows show the neural cell-specific neurite outgrowth. Scale bar, 200 μm. (f) Gene expression analysis of Nodal. After attachment at day 1, the anterior cells were treated with DMSO (A-DMSO) or TSA (A-TSA) for 24 h, and harvested for the messenger RNA expression analysis. Relative expression level was normalized by gapdh. (g) Gene expression analysis of the day 5 cells described in e. (Sox1, n=8; Pax6, n=5; Mixl1, n=6; Flk1, n=5). Relative expression level was normalized by gapdh. Representative data of qPCR (d,f) were shown from three independent experiments. Data in b,d,f,g represent mean±s.d. *P<0.05, Student’s t test.