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Fig. 9. Assays on the transcriptional activity of Klf4. (A) Construction of fusion constructs of Klf4. (B) The effect of the injection of 150 pg of VP16-Klf4(DBD) or eve-Klf4(DBD) mRNA on gene expression. Embryos at stage 10.5 were used for detection of Dkk1 and embryos at stage 26 were used for detection of XAG2. In the panels for Dkk1 expression, the middle one was lateral view and the other two were vegetal view. In the panels for XAG2 expression, the first two from the left are ventral view and the third is anterior view. (C) Quantification of embryos with normal or altered gene expression observed in B in triplicate. (D) The effect of fusion constructs on ARELuc using luciferase assays in HEK 293T cells. (E,F) The effect of fusion constructs on Mix2 promoter/luciferase reporters in embryos. VP16-Klf4(DBD) or eve-Klf4(DBD) mRNA (150 pg), and 40 pg of each reporter plasmid were injected. Embryos were collected at stage 10 for luciferase activity measurement. Error bars represent s.d. in eight (D) or six (E,F) experiments. |
