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. 2014 Mar 26;5(6):1666–1682. doi: 10.18632/oncotarget.1850

Figure 1. Characterization of NWD1.

Figure 1

(A) Domain structure of NWD1. The protein contains a centrally located NACHT region and 13 WD40 repeats (WD1-13). Domain positions are indicated (bottom). (B) Tissue-specific expression of NWD1. A representative graph illustrating the relative NWD1 expression in 48 independent human tissues is shown. First-strand cDNA derived from each tissue sample was normalized according to GAPDH expression (Human Major Tissue qPCR Array, HMRT102, Origene). (C) Subcellular localization of NWD1. HEK 293T cells cultured in a 10-centimeter plate to 90% cell confluency were transfected by calcium phosphate with 10 μg of FLAG-tagged NWD1 expression vector. After 48 hours, cells were fractionated. A total of 50 μg of respective nuclear and cytosolic extract fractions were resolved by SDS-PAGE, and further analyzed by immunoblotting using antibodies specific for α-tubulin (cytosolic marker), PARP (nuclear marker) and FLAG-epitope tag (for detection of ectopic NWD1).