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. Author manuscript; available in PMC: 2021 Mar 29.
Published in final edited form as: Dev Cell. 2020 Dec 14;56(3):277–291.e6. doi: 10.1016/j.devcel.2020.11.017

Figure 2. ESCs Lose Their Stem Cell Phenotypes When Cultured in N2B27 Medium.

Figure 2.

(A) Cell growth assays were performed on ESCs maintained in 2i+LIF or N2B27 medium.*

(B) Cell death assays were performed on ESCs maintained in 2i+LIF or N2B27 medium.*

(C) ESCs cultured in 2i+LIF or N2B27 medium were immunoblotted for Oct3/4, Nanog, the trimethylation of lysine 27 in histone H3 (H3K27me3), and vinculin as the loading control.*

(D) Sphere formation assays were performed on ESCs maintained in 2i+LIF or N2B27 medium. See corresponding images in Figure S2B.*

(E) AP activity assays were performed on ESCs maintained in 2i+LIF or N2B27 medium. See corresponding images in Figure S2C.*

*The data shown in (A)–(E) are presented as mean ± SD. All experiments were performed at least three independent times, and statistical significance was determined using Student’s t test; ****p < 0.0001; ***p < 0.001; and **p < 0.01. See also Figure S2.