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. 2010 Nov 1;24(21):2408–2419. doi: 10.1101/gad.1987810

Figure 5.

Figure 5.

Increased Δ40p53 inhibits ESC differentiation. (A) Larger size of EBs with increased Δ40p53. Quantitative analysis of mean diameters of ICR and p44Tg EBs ± SEM after 4 d of EB culture. (***) P < 0.001 (two-tailed Student's t-test). (B) Improved survival of EBs with increased Δ40p53 EB viability was calculated based on the percentage of all wells plated containing at least one EB after 4 d of culture. Mean survival rates ± SEM are displayed. (***) P < 0.001 (two-tailed Student's t-test). (C–E) Impaired response to differentiation conditions in cells with increased Δ40p53. (C) Proliferation of cells during monolayer differentiation. Data are displayed as the fold change in cell number based on the number of ESCs plated on day 1. (Blue) p44Tg; (red) ICR. (D) Phenotype of cells during monolayer differentiation. Photomicrographs of ICR and p44Tg ESCs at days 0, 7, and 14 of monolayer differentiation. (Top panels) Phase contrast. (Bottom panels) Immunofluorescence with an antibody against SSEA-1. Original magnification, 40×. (E, top panel) Western blot analysis of Nanog and Oct4 expression in ICR and p44Tg ESCs and day 5 EBs. Protein quantitation in ESCs (black bars) and EBs (gray bars), normalized to actin, is displayed below. The Oct4 antibody detects a doublet at 44/42 kDa; both bands of the doublet were measured for the quantitation.