Fig. 7: ITZ inhibits the proliferation of human ES cell lines in culture and suppresses the growth of ES xenografts in mice.

a Viability TC-71 cells treated with DMSO or ITZ for 72 h. Data are presented as mean ± SD (n = 3 biologically independent experiments). P-values were calculated using one-way ANOVA with post-hoc Dunnett’s multiple comparisons test. b Representative immunoblot showing PARP levels in TC-71 cells treated with DMSO or ITZ for 48 h. Cleaved PARP is indicated by the arrow. c Viability of TC-71 cells expressing vector, C1GALT1 or EWSR1::FLI1 and treated with DMSO or ITZ for 72 h. Data are presented as mean ± SD (n = 3 biologically independent experiments). P-values were calculated at 100 nM ITZ using two-way ANOVA with post-hoc Tukey’s multiple comparisons test. d Tumor formation. TC-71 cells, either parental or expressing vector or EWSR1::FLI1, were injected subcutaneously into NSG mice and when tumors reached ~100 mm³ (denoted as day 0), mice were treated daily with vehicle or ITZ (100 mg/kg), and tumor dimensions were monitored every 3 days. Data are presented as mean ± SD (n = 6 mice per group). P-values were calculated on day 18 using two-way ANOVA with post-hoc Tukey’s multiple comparisons tests. e Representative IHC images showing Ki67, C1GALT1, SMO, GLI1 and FLI1 staining in TC-71 xenografts at day 18. Scale bar, 50 µm. f Tumor formation. TC-71 cells expressing a NS or C1GALT1 shRNA were injected subcutaneously into NSG mice, and when tumors reached ~100 mm³ (day 6 for NS shRNA-expressing tumors or day 9 for C1GALT1 shRNA-expressing tumors), mice were treated daily with vehicle or ITZ. Data are presented as mean ± SD (n = 6 mice per group). P-values were calculated following 15 days of treatment using mixed-effect analysis with post-hoc Tukey’s multiple comparisons tests. g Representative IHC images showing C1GALT1, SMO, GLI1, and FLI1 staining in tumors derived from mice in (f). Scale bar, 50 µm. Source data are provided as a Source Data file.