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. 2017 Dec 8;19(2):337–350. doi: 10.15252/embr.201744777

Figure 2. TRIP6 competes with MOB1 for binding to LATS2.

Figure 2

  1. LATS2‐GFP and Myc‐MOB1A were overexpressed in HEK293 cells with or without co‐overexpression of full‐length FLAG‐TRIP6 and FLAG‐TRIP6 1–277. Myc‐MOB1A was immunoprecipitated using anti‐Myc antibodies, and immune complexes were assayed for Myc‐MOB1A and LATS2‐GFP levels. Levels of FLAG‐TRIP6, FLAG‐TRIP6 1–277, Myc‐MOB1A, and LATS2‐GFP in the lysate are also shown. The levels of LATS2‐GFP in immune complexes relative to the level of Myc‐MOB1A are shown in the graph (mean ± SD; n = 3; **P ≤ 0.01, t‐test).
  2. Competitive binding experiments were done using purified recombinant MBP‐LATS2, GST‐TRIP6, and 6His‐MOB1A. MBP‐LATS2 bound to maltose beads was incubated with GST‐TRIP6 with or without increasing amounts of 6His‐MOB1A, and the levels of each protein bound to MBP‐LATS2 on the beads at the end of the experiment were determined by Western blotting. The levels of input proteins are shown (lysate). The binding of 6His‐MOB1A and GST‐TRIP6 to MBP alone, and the use BSA as a competitor instead of 6His‐MOB1A are shown as controls. The numbers at the bottom are referred to in the text.