Fig. 5.

EphA7 is a downstream target of Zfp422 and rescues the differentiation of Zfp422-KO C2C12 myoblasts. a C2C12 cells were transfected with 3.1-Zfp422-Myc vector to introduce exogenous Myc-tagged Zfp422 expression, 3.1-GFP-Myc vector acted as a control. Then ChIP-seq was performed using Myc antibody. PCR validated the 169bp sequence bound by Zfp422 from the ChIP-seq results. b, c EphA7, EphA4 and Pald1 mRNA expression in Zfp422-KO C2C12 cells. d EphA7 protein expression in Zfp422-KO and control C2C12 cells. e Schematic of reconstructed pGL3-baisc luciferase reporter vector. f C2C12 cells were transiently transfected with Zfp422 expression vector or empty vector, together with other indicated luciferase reporter vectors, then measured dual luciferase activity 48 h later. g Zfp422-KO and control C2C12 cells were transiently transfected with the indicated luciferase reporter vectors, then harvested and measured dual luciferase activity 48 h later. h, i EphA7 mRNA expression (h) and protein level (i) in Zfp422-KO C2C12 cells after transfection with 3.1-EphA7 vector. j The expression changes of genes related to myoblast differentiation after EphA7 overexpression in Zfp422-KO C2C12 cells. k Immunostaining for MyHC at DM3d after control or EphA7-expression vector transfection in Zfp422-KO cells. Differentiation index was counted. Data are presented as mean ± SD, n = 3 for each group. *p < 0.05, **p < 0.01, ***p < 0.001. n.s., no significance