Extended Data Figure 2: KBTBD8 controls neural crest formation.
From: Cell-fate determination by ubiquitin-dependent regulation of translation
a, Stable depletion of KBTBD8 from H1 hESCs, as determined by western blot analysis. b, KBTBD8 depletion does not significantly change the cell cycle profile of hESCs, as determined by propidium iodide staining and FACS. c, Control or KBTBD8-depleted hESCs were counted at indicated times after seeding (mean of 3 biological replicates, ±s.d.). d, KBTBD8 depletion does not induce apoptosis in hESCs, as shown by immunostaining against cleaved caspase 3 (red) or DNA (blue) (200 cells per condition; scale bar, 10 μm). e, KBTBD8 depletion does not affect the gene expression profile of hESCs, as determined by microarray analysis (genes > 2.5-fold change, n > 30,000; mean of 3 biological replicates, ANOVA P-value <0.05). f, Loss of KBTBD8 causes a decrease in the expression of neural crest cell markers during EB formation, as shown by comparative microarray analysis (genes > 2.5-fold change, n > 30,000; mean of 3 biological replicates, ANOVA P-value <0.05). g, mRNA levels of pluripotency and differentiation markers in EBs stably expressing control or KBTBD8 shRNAs were measured by qRT–PCR (3 technical replicates ± s.e.m.).
