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. 2010 May 19;24(7):1478–1497. doi: 10.1210/me.2010-0029

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Copyright © 2010 by The Endocrine Society

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HIPK3 interacts with Runx2 and with MINT MID and colocalizes with MINT in the nucleus. A, In vitro synthesized and unlabeled V5-tagged-HIPK3 was incubated with radioactively labeled OPN or Runx2 and immunoprecipitated with mouse anti-V5 antibody. Note that radioactive Runx2 (lanes 2 and 6–8), but not OPN (lanes 1 and 3–5), is coprecipitated by pull-down of V5 HIPK3. B, CMV-MINT was transfected into C3H10T1/2 cells and localization compared with that of endogenous HIPK3 (upper row) or coexpressed EYFP-HIPK3 (lower row) by confocal microscopy. The two proteins extensively colocalize in the nucleus and exclude the nucleoli. C, In vitro synthesized and unlabeled V5-tagged-HIPK3 was incubated with radioactively labeled OPN or T7-tagged MID MINT and immunoprecipitated with mouse anti-V5 antibody. Note that radioactive MID MINT (lanes 2, 5, and 6), but not OPN (lanes 1, 3, and 4), is coprecipitated by pull-down of V5 HIPK3. IP, Immunoprecipitated.