FIG. 10.

TERT morpholinos induce spontaneous hESC differentiation that appears to be lineage biased. hESCs grown in feeder-free conditions were assayed for lineage-specific expression after exposure to steric-blocking morpholino oligonucleotides (MOs) specific for alternative splice sites of TERT pre-mRNA. hESCs were passaged in the presence of MO to facilitate uptake. Controls were hESCs grown in medium containing no basic fibroblast growth factor (−bFGF) to induce spontaneous differentiation. Real-time RT-qPCR analysis of hESCs treated with 10 μM of either Δα MO, Δβ MO, or standard control MO for 3 days. Graphs show real-time RT-PCR analysis for various markers: pluripotency (Oct3/4 and Nanog), ectoderm [neurogenic differentiation factor 1 (NeuroD1) and paired box 6 (PAX6)], endoderm [GATA6, GATA4, and alpha-fetoprotein (AFP)], and mesoderm (T and Brachyury). All samples were normalized to the large ribosomal protein (RPLPO) as an internal control, and fold changes calculated using the delta-delta-CT method. Error bars represent SE, n=3. Letters above histogram bars indicate significant differences (P<0.05).