Skip to main content
. 2012 Feb 28;287(16):12994–13004. doi: 10.1074/jbc.M111.323105

FIGURE 3.

FIGURE 3.

JMJD5 repressed transcriptional activity of NFATc1 by reducing its protein level. A, HEK293T cells were transfected with Myc-NFATc1 together with FLAG-mJMJD5-WT or FLAG-mJMJD5-K334A. After 48 h of transfection, total cell lysates were prepared and subjected to immunoprecipitation (IP) with anti-FLAG M2-agarose. Bound proteins were eluted by FLAG peptides and detected by immunoblotting with the indicated antibodies. An asterisk indicates a nonspecific band. B and C, NFATc1-RE gene was established with the indicated region of the Acp5 gene promoter (B, left). HEK293T cells were transfected with NFATc1-RE together with FLAG-NFATc1 and FLAG-mJMJD5-WT or FLAG-mJMJD5-K334A. After 36 h of transfection, cells were analyzed with a luciferase assay (B, right) or by immunoblotting (C). The error bars represent S.D. values from at least triplicate experiments. GAPDH was used as loading control for immunoblotting. An asterisk indicates a nonspecific band. The signals obtained from the immunoblotting were quantified with the ImageJ program. TSS, transcription start site.