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. Author manuscript; available in PMC: 2017 Feb 1.
Published in final edited form as: Nat Cell Biol. 2016 Aug 1;18(9):917–929. doi: 10.1038/ncb3394

Figure 6. Mapping of Snail/Slug-YAP/TAZ Binding Interactions and Regulation of Transcriptional Activity.

Figure 6

(a) 8XGTIIC luciferase activity was determined in Cos-1 cells co-transfected with YAP, TAZ, Snail or Slug (mean ± s.d., n=3 independent experiments).

(b) Snail/Slug binding to YAP/Tead4 complexes. Epitope-tagged Snail, Slug, YAP and Tead4 were co-transfected into Cos-1 cells, lysates immunoprecipitated with anti-Myc antibody (Tead4 tagged with Myc) and immunoblotted. Results are representative of 3 experiments performed.

(c) 6XOSE luciferase activity was monitored in Cos-1 cells co-transfected with Runx2, Snail, Slug, YAP or TAZ (mean ± s.d., n=3 independent experiments).

(d) Snail/Slug incorporation into TAZ/Runx2 complexes. Epitope-tagged Snail, Slug, YAP or Runx2 were co-transfected into Cos-1 cells, lysates immunoprecipitated with anti-HA antibody (Runx2 tagged with HA) and samples immunoblotted. Results are representative of 3 experiments performed.

(e) Schematic of HA-tagged Snail mutants.

(f) YAP interacts with the SNAG domain of Snail. Cos-1 cells were co-transfected with the indicated Snail-HA or mutant constructs in tandem with Flag-YAP. Cell lysates were subjected to anti-HA immunoprecipitation, and YAP in the complexes determined by anti-Flag immunoblotting. Results are representative of 3 experiments performed.

(g) TAZ interacts with the SNAG domain of Snail. Cos-1 cells were co-transfected with the indicated Snail-HA or mutant constructs in tandem with Flag-TAZ. Cell lysates were subjected to anti-Flag immunoprecipitation, and wild-type Snail or Snail mutants in the complexes assessed following anti-HA immunoblotting. Results are representative of 3 experiments performed.

(h) 8XGTIIC luciferase activity was measured in Cos-1 cells following co-transfection with YAP, TAZ, Snail or Snail mutants (mean ± s.d., n=3 independent experiments).

(i) 8XGTIIC luciferase activity was determined in Cos-1 cells co-transfected with YAP, TAZ, Slug or Slug mutants (mean ± s.d., n=3 independent experiments).

(j) 6XOSE luciferase activity was determined in Cos-1 cells co-transfected with Runx2, TAZ, Snail or Snail mutants (mean ± s.d., n=3 independent experiments).

(k) 6XOSE luciferase activity was monitored in Cos-1 cells co-transfected with Runx2, TAZ, Slug or Slug mutants (mean ± s.d., n=3 independent experiments).