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. 2015 Jun 9;5:10449. doi: 10.1038/srep10449

Figure 4. PLK1 restricts NDR1 kinase activity through interfering with activator Mob1-binding (a).

Figure 4

Pull-down assay of MBP-NDR1 wild-type or mutants (WT, 3A and 3E) using GST- or GST-Mob1A-bound agarose beads. The pulled-down MBP-NDR1 bands were quantified by Image J software and normalized. (b) Co-immunoprecipitation of Flag-NDR1 (WT, 3A and 3E) with GFP-Mob1A and mCherry-Mob2. Lysates of asynchronized 293 T cells expressing indicated plasmids were subjected to immunoprecipitation with Flag antibody. The precipitates (IP) and cell lysates were analyzed by immunoblot with antibodies against Flag, GFP and Cherry, respectively. (c) Co-immunoprecipitation of LAP-NDR1 with endogenous Mob1 and Mob2. LAP-NDR1 stable expressing HeLa cells were treated as in Fig. 3h. The lysates were subjected to immunoprecipitation with GFP-Trap agarose beads. The precipitates (IP) and cell lysates were analyzed by immunoblot with antibodies against GFP, actin, Mob1 and Mob2. Cropped blots from same gel.