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. 2017 Sep 4;6(10):1528–1540. doi: 10.1242/bio.026153

Fig. 8.

Fig. 8.

Vgll3 interacts with ets1 and activates a tead-dependent luciferase reporter gene. (A) Pax3 expression in embryos injected with vgll3 or ets1 mRNA. Ets1 overexpression induces ectopic pax3 expression as well as co-expression of ets1 and vgll3. Asterisks indicate the injected side. (B) Embryos were injected with myc-ets1 or HA-vgll3 mRNAs. Immunoprecipitation with myc (IP myc) or HA antibodies (IP HA) was followed by IB with antibodies. (C) Schematic representation of the pGL284LUC reporter plasmid used in transfection assay. The different cis-sequences are depicted together with the MCAT cis-sequence (black box). (D) Left panel: ets1, tead1 and vgll3 are expressed in HEK293 cells when analysed by RT-PCR. (−), no cDNA. Right panel: HEK293 cells were transfected with HA-vgll3, HA-V3ΔTDU, myc-tead1 or myc-ets1 plasmids and similar amounts of proteins were checked by IB with myc and HA antibodies. Gapdh and erk2 are used as control. (E) HEK293 cells were co-transfected with pGL284LUC and HA-vgll3, HA-V3ΔTDU, myc-tead1 and myc-ets1 plasmids. Relative luciferase activity is expressed as fold change in luciferase activity compared to tead1 or ets1 alone. Data are mean±s.e.m. from three independent experiments carried out in duplicate. *P<0.05; Student's t-test.