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Expression and subcellular localization of MEOX1 and MEOX2 proteins.

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posted on 20.12.2011 by Josette M. Douville, David Y. C. Cheung, Krista L. Herbert, Teri Moffatt, Jeffrey T. Wigle

A) Schematic representation of the MEOX1 and MEOX2 protein constructs used in this paper. The alignment lists the percentage of similar and identical amino acids conserved between human MEOX1 and MEOX2 in the different functional protein domains. B) Representative western blot displaying the relative level of MEOX1 (MX1), MEOX2 (MX2), DNA binding deficient MEOX1Q220E (Q220E), DNA binding deficient MEOX2Q235E (Q235E) and homeodomain deleted MEOX2K195_K245del (K195_K245del) protein expression in HEK293 cells 48 hours after transfection. The N-terminally tagged MEOX proteins were detected using an anti-FLAG antibody and α-tubulin was used as a loading control. The empty expression vector was used as a negative control (Ctrl). C) A representative western blot demonstrating the subcellular localization of different MEOX proteins in HEK293 cells, 48 hours after transfection. α-tubulin was used as a cytoplasmic (C) marker and lamin A/C was used as nuclear (N) marker. D) Representative fluorescent immunocytochemistry showing the localization and level of expression of the MEOX proteins in HUVECs 48 hours after adenoviral transduction at a multiplicity of infection of 250. The N-terminally tagged MEOX proteins were detected using an anti-FLAG antibody (green) and nuclei were stained with propidium iodide (red). Enhanced green fluorescent protein (EGFP) was used as a control for adenoviral infection. Arrows indicate cytoplasmic staining and arrowheads indicate punctate nuclear aggregates. Scale bar represents 20 µm.