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. 2008 Jun 16;181(6):959–972. doi: 10.1083/jcb.200711044

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© 2008 Li et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 3. — CHIP inhibits Runx2-induced transcriptional activity. (A) Overexpression of CHIP inhibits Runx2-induced transcriptional activity. Luciferase assays were performed using NIH3T3 or COS7 cells transfected with the indicated expression vectors along with a Runx2-responsive reporter construct, 6xOSE2-OC/pGL3 and pRL-TK (internal control). Values were normalized using the internal control and presented relative to basal activity (NIH3T3, first bar; COS7, fifth bar) with the mean from three independent repeats. (B) Mutants of CHIP fail to inhibit Runx2-induced transcriptional activity. CHIP, CHIP mutants, and Flag-Smurf1 were compared for the inhibitory effect on Runx2 activity in NIH3T3 cells. (C) Knocking down CHIP by siRNA facilitates Runx2-mediated transcription in UMR106 cells. Luciferase assays were performed in UMR106 cells transfected with Runx2 in the presence of CHIP, EGFP RNAi, or CHIP RNAi. (D) CHIP did not affect the transcriptional activity of STAT3. NIH3T3 cells transfected with STAT3 in the presence or absence of CHIP along with pGL3–acute phase response element and treated with 10 ng/ml interleukin-6. All of the data are presented as mean with SEM (error bars) from three independent experiments. The same letter represents a significant difference within this group in a meaningful comparison (P < 0.05).