Figure 3. Pou5f1 and Cebpβ directly regulate aid, mbd4 and gadd45α expression in apc^mcr embryos.

(A) Whole mount in situ staining for aldh1a2 in apc^mcr and apc^wt at 72hpf. (B-C) Whole mount in situ staining for fabp2 in apc^mcr and apc^wt embryos (B) or in control morphant and rdh1l morphant embryos (C) (at 72hpf) treated with vehicle or RAL (2μM). (D) RT-PCR for Pou5f1 (Oct4) and Cebpβ in apc^mcr and apc^wt treated with DMSO or ATRA (1μM). The Y-axis shows fold induction normalized to 28S and wild type DMSO treated sample. (E-F) Whole mount in situ staining for aid, gadd45α and mbd4 in apc^mcr and apc^wt injected with control, Pou2f1a, pou5f1 or cebpβ Mo (E) or in wild type embryos injected with control, Pou2f1, pou5f1 or cebpβ expression plasmid (F). (G) Graph showing fold enrichment near the aid or gadd45α TSS (containing overlapping Oct and Cebp binding sites) for Cebpβ and Pou5f1 in embryos injected with V5-Cebpβ (along with Pou5f1 Mo, 80pg), V5-Pou5f1 or FLAG-Pou2f1 expressing plasmids. ChIP was performed with antibodies against the tags. Normalization control primers are located ~3kb upstream (region without Cebpβ sites) of TSS of Gadd45α gene. Error bars indicate +/− SD. See also Figure S3.