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. 2015 Jul 7;290(33):20273–20283. doi: 10.1074/jbc.M115.646554

FIGURE 2.

FIGURE 2.

Jmjd6 interacts with Tcf7l1. A, co-IP detection of the interaction between overexpressed Jmjd6 and Tcf7l1 in HEK293T cells. IB, immunoblot. MT, myc tag. B, overexpressed Tcf7l1 precipitated endogenous JMJD6 in HEK293T cells. C, nuclear colocalization of Jmjd6 and Tcf7l1 in HEK293T cells as detected by immunofluorescence staining. DAPI staining reveals nuclei. D and E, test of the knockdown efficiency of miJMJD6-1. Transfection of the plasmid for miJMJD6-1 did not cause a significant reduction in both the transcript (D) and protein (E) levels of JMJD6 in HEK293T cells. RT−, transcription without reverse transcriptase. F and G, test of the knockdown efficiency of miJMJD6-2 and the effect of JMJD6 knockdown on the transcription of TCF7l1 and β-Catenin (β-Cat). Transfection of the plasmid for miJMJD6-2 resulted in a significant decrease in both the transcript (F) and protein (G) levels of JMJD6. Meanwhile, the transcription of TCF7L1 and β-Catenin was not affected in response to efficient JMJD6 knockdown. GAPDH was used as a loading control for RT-PCR detection of the JMJD6 transcript in D and F, and β-Actin was used as a loading control for the detection of JMJD6 protein with immunoblotting in E and G. H, detection of the effect of endogenous JMJD6 knockdown on the interaction between exogenous Jmjd6 and Tcf7l1.