Figure 2: HDACi inhibits mESC neural commitment and promotes mesendoderm differentiation.

(a) Statistical analysis of Sox1–GFP⁺ cells at day 6 (n=4). Day 2 cells were treated with DMSO or TSA (10 nM, 20 nM, 50 nM) for 24 h. (b) Western blotting analysis of H3ac and H3K9ac levels during mESC neural differentiation. Day 2 cells were treated with DMSO or TSA for 24 h, then removed and harvested from day 2 to day 6. (c) Gene expression analysis of day 6 cells described in b. Relative expression level was normalized by gapdh. (d) Sox1–GFP⁺, Pax6, Flk1 and Gata6 immunocytochemistry of the day 6 cells described in b. Scale bar, 75 μm. (e) Statistical analysis of Sox1⁺, Pax6⁺, Flk1⁺ and Gata6⁺ cells in d (n=4). (f) Statistical analysis of Sox1–GFP⁺ cells at day 6 (n=4). DMSO or TSA was added at different time point from day 2 to day 5 for 24 h. (g) Gene expression analysis of day 6 cells described in f. Relative expression level was normalized by gapdh. Unless indicated, the data were shown from at least three independent experiments. Data in c,e–g represent mean±s.d. *P<0.05, **P<0.01, Student’s t-test.