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. 2017 Jun 1;9(1):92–107. doi: 10.1016/j.stemcr.2017.04.032

Figure 2.

Figure 2

THAP1 Is Not Required for ESC Maintenance

(A) Immunofluorescence analysis of POU5F1 (upper panel) SSEA1 (middle panel), and alkaline phosphatase (AP) staining (lower panel) in wild-type (WT), Thap1C54Y/C54Y (KI), and Thap1ΔExon2 (KO) ESCs.

(B) Proliferation rate of wild-type (WT), Thap1C54Y/C54Y (KI), and Thap1ΔExon2 (KO) ESCs relative to day 0 and measured by counting the cells at the indicated time points. Two-way repeated-measure (RM) ANOVA was performed revealing an interaction effect between days and genotype (F(8,36) = 4.288, p = 0.0011). In addition, significant differences were observed by day (F(4,36) = 139.4, p < 0.0001), while marginal differences were observed by genotype (F(2,9) = 3.685, p = 0.0677). The Holm-Sidak multiple comparisons test was performed post hoc, revealing significant differences between the genotypes. Data are presented as mean ± SEM of four independent experiments. ∗p < 0.05, ∗∗∗p < 0.005, ∗∗∗∗p < 0.001.

(C) Percentage of live (Annexin V−) and dead cells (Annexin V+) in wild-type (WT), Thap1C54Y/C54Y (KI), and Thap1ΔExon2 (KO) ESCs relative to WT. Two-way RM ANOVA was performed revealing an interaction effect (F(2,24) = 21.29, p < 0.0001). In addition, significant differences were observed by apoptotic marker (F(4,36) = 139.4, p < 0.0001) and by genotype (F(2,24) = 4.205, p = 0.0272). The Holm-Sidak multiple comparisons test was performed post hoc, revealing significant differences between the genotypes. Data are presented as mean ± SEM of five independent experiments. ∗∗∗∗p < 0.001.

(D) Cell-cycle distribution of wild-type (WT), Thap1C54Y/C54Y (KI), and Thap1ΔExon2 (KO) ESCs relative examined by DNA content index. Two-way RM ANOVA was performed, revealing an interaction effect (F(4,48) = 14.61, p < 0.0001). In addition, significant differences were observed by cell-cycle phase (F(2,48) = 21.52, p < 0.0001), but not by genotype. The Holm-Sidak multiple comparisons test was performed post hoc, revealing significant differences between the genotypes for S and G2/M cell-cycle phases. Data are presented as mean ± SEM of four independent experiments. ∗∗p < 0.01, ∗∗∗∗p < 0.001.