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. Author manuscript; available in PMC: 2016 Dec 7.
Published in final edited form as: Oncogene. 2016 Jun 6;36(1):97–109. doi: 10.1038/onc.2016.179

Figure 6. CASZ1b contains nuclear export signal (NES) at the N-terminus.

Figure 6

(A) The alignment of human CASZ1 (hcas) and zebrafish CASZ1 (zcas) at the N-terminal region that contains a putative NES with 5 conserved hydrophobic AAs (underlined in red) between AA175 to AA191. (B) The NES-NLS2-GFP and NES mutant-NLS2-GFP constructs. CASZ1b AA170–249 region that contains both putative NES and NLS2 was fused to GFP. NESm-NLS2-GFP is a NES mutant constructed by replacing the 5 conservative hydrophobic AAs within putative NES and one hydrophobic AA close to NES with alanine (highlighted in red). (C) 293T cells and (D) AS cells were transiently transfected with NLS2-GFP, NES-NLS2-GFP and NESm-NLS2-GFP constructs, chromatin was stained with DAPI. A minimum of 300 cells were screened for each group from at least 3 different fields and scored for nuclear and cytoplasmic (Nuc & Cyt) or nuclear only (Nuc) staining (data are shown as means ± S.D., *: p<0.001). None of the cells have cytoplasmic only GFP signal, thus Cyt was not shown in the graph.