Figure 1. Undifferentiated hESCs express only a subset of TFIID TAFs.
(A) Immunoblot analysis showing TAF levels in HeLa cells and H9 hESCs. β-actin (ACTB) was monitored as a loading control. (B) qRT-PCR analysis monitoring TAF expression in H9 cells relative to HeLa cells. A ratio of 1 (indicated by the red dotted line) indicates no difference in expression. Data are represented as mean ± SEM. (C) Immunoblot analysis showing levels of GTFs in HeLa cells and H9 hESCs. α-tubulin (TUBA) was monitored as a loading control. (D) Immunoblots showing TAF and TBP levels in H9 hESCs following induction of differentiation by retinoic acid treatment for 0, 3 or 6 days. OCT4 and NES were monitored as controls.
Figure 1—figure supplement 1. Confirmation of specificity of TAF antibodies by RNAi-mediated knockdown in H9 hESCs.
Immunoblot analysis showing TAF levels in H9 hESCs 48 hr after transfection with a control luciferase (Luc) or TAF siRNA. β-actin (ACTB) was monitored as a loading control.
Figure 1—figure supplement 2. Confirmation of specificity of TAF antibodies by RNAi-mediated knockdown in HeLa cells.
Immunoblot analysis showing TAF levels in HeLa cells 48 hr after transfection with a control luciferase (Luc) or TAF siRNA. β-actin (ACTB) was monitored as a loading control.
Figure 1—figure supplement 3. TAF expression levels in H1 hESCs.
Immunoblot analysis showing TAF levels in H1 hESCs and, as a comparison, H9 hESCs and HeLa cells. β-actin (ACTB) was monitored as a loading control. The results demonstrate an identical pattern of TAF expression in both H9 and H1 hESCs.