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. Author manuscript; available in PMC: 2010 Jun 11.
Published in final edited form as: Neuron. 2009 Jun 11;62(5):641–654. doi: 10.1016/j.neuron.2009.04.025

Figure 2. CBP is a key effector for RA-signal to promote Ngn2 function.

Figure 2

(A) RA enhances in vivo association between Ngn2 and CBP in CoIP experiments, as tested with HEK293 cells transfected with HA-Ngn2. (B) CoIP assays using IP with anti-HA antibody followed by immunoblotting with anti-CBP antibody in HEK293 cells transfected with HA-Ngn2 and CBP along with either RAR or RARΔAF2. RAR expression further strengthens the association of Ngn2 and CBP, while RARΔAF2 attenuates this interaction. (C, D) Quantitative-RT-PCR for neurofilament M (NF-M) (C) and NeuroD (D) in P19 cells treated as indicated. (E) Immunohistochemical analysis of neuronal differentiation (TuJ+-cells) in HH stage 22 chick embryos electroporated with the indicated constructs on the left. Cells coexpressing Ngn2 and E1A fail to differentiate to TuJ+-neurons, suggesting that Ngn2-mediated neurogenesis is suppressed by E1A. In contrast, E1AΔN does not block neurogenesis in the medial zone of the chick neural tube (data not shown). (F) Quantitative analysis of neuronal differentiation in the chick neural tube. (C, D, F) The error bars represent the standard deviation.