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. Author manuscript; available in PMC: 2012 Aug 4.
Published in final edited form as: FEBS Lett. 2011 Jun 26;585(15):2410–2418. doi: 10.1016/j.febslet.2011.06.024

Figure 2. Direct binding and intracellular co-localization of NELL-1 and APR3.

Figure 2

A. APR3 and NELL-1 expression plasmids or empty vectors were cotransfected into Cos-7 cells. APR3 protein with FLAG tag and NELL-1 protein were pulled down respectively by anti-FLAG affinity gel and Dynabeads Protein G binding with anti-NELL1 antibody. Western blot was performed after pull down assay. NELL-1 was detected in the APR3 pull down sample, but not in the vector sample. APR3 was detected in the NELL-1 pull down sample, but not in the vector sample.

B-C. APR3 expression plasmid with red fluorescence tag and NELL-1 expression plasmid with green fluorescence tag were cotransfected into Saos2 (B) and U2OS (C) cells. Samples were observed by confocal microscope after 48 hours of expression. NELL-1 was localized in the cytoplasm of single positive cells (arrowheads) while APR3 and NELL-1 were found to be co-localized in the perinuclear region and on the nuclear envelope of double positive cells as indicated by arrows.