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. 2005 Nov 30;33(21):6782–6794. doi: 10.1093/nar/gki979

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© The Author 2005. Published by Oxford University Press. All rights reserved

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Characterization of HOXA13-FLAG/EGFP or HOXD13-FLAG/EGFP expressing cells. (A) Western blot using HOX-specific antibodies demonstrating HOXA13 expression and anti-FLAG immunoprecipitation from HOXA13-FLAG cell line and absent HOXA13 expression in the HOX (−) cell line (1); HOXD13-FLAG expression and anti-FLAG immunoprecipitation from HOXD13-FLAG cell line but absent HOXD13 expression in the HOX (−) cell line (2). (B) Immunocytochemistry using Hox specific antibodies and DAPI staining demonstrate expression and nuclear localization of HOXA13 or HOXD13 in their respective cell lines. (C) Input RNA using serial dilutions ranging from 156 pg to 10 ng was used in semi-quantitative RT–PCR to look for expression changes of four reported targets. Fhl1 (+6-fold), Enpp2 (+18.8-fold) and M32486 (+2.3) are upregulated in the HOXA13-FLAG (A) and HOXD13-FLAG (D) cell lines and Ngef (−2.4) is downregulated compared to HOX (−). Water (W) was used as a PCR control. Ppic was a loading control and was shown to be unchanged in the HOXA13-FLAG and HOXD13-FLAG cell lines.