Fig. 5.

E2A directly regulates transcription factors associate with the development of cardiac conduction cells and multiple signaling pathways

(a) Expression of transcription factors SHOX2, TBX3, TBX5 and TBX18 in day3, 7, and 15 CMs post differentiation. (b) mRNA levels of SHOX2, TBX3, TBX5 and TBX18 in day30 CMs. Data were normalized to GAPDH level and presented as mean ± SEM. * P value < 0.05, ** P value < 0.01, *** P value < 0.01, # P value < 0.0001(Two-tailed Student's t-test) (c) E2A binding profile from http://jaspar.genereg.net/. (d) E2A binding sites were predicted in chromatin open regions of SHOX2, TBX3, TBX5 and TBX18 in hESCs (CistromeDB:79373), human CPC (CistromeDB:80590), human CM (CistromeDB:80589), and fetal heart tissue (CistromeDB:40936), arrows indicate transcription start site (TSS) of genes, E-box elements are marked by red lines below and corresponding sequences are listed in the gray boxes. (e) Dual-Luciferase reporter assay in 293T showed that E2A directly regulated expression of SHOX2, TBX3, and TBX5, but not TBX18. Data were presented as mean ± SEM. # P value < 0.0001(Two-tailed Student's t-test) (f) KEGG analyses of up-regulated signal pathway in day0,3,7,15 E2A deficient cells. (g) Top 10 predicated upstream regulators.