Journal: Nature cell biology

Article Title: Snail/Slug-YAP/TAZ Complexes Control Skeletal Stem Cell Self-Renewal and Differentiation

doi: 10.1038/ncb3394

Figure Lengend Snippet: (a) Western blot of p-YAP, p-TAZ, p-Lats1 and p-Mst1 in SSCs isolated from Snail f/f /Slug +/+ and Snail f/f /Slug −/− mice and transduced with adeno-GFP or Cre. Results are representative of 3 experiments performed. (b) Association of YAP/TAZ with Snail was detected by immunoprecipitation in Cos-1 cells transfected with Flag-YAP, Flag-TAZ and Snail-HA, respectively, as indicated. Results are representative of 3 experiments performed. (c) Association of YAP/TAZ with Slug was detected by immunoprecipitation in Cos-1 cells transfected with Flag-YAP, Flag-TAZ and Slug-Myc, respectively, as indicated. Results are representative of 3 experiments performed. (d) Associations between endogenous YAP/TAZ and Snail/Slug were detected in SSCs cultured in the absence or presence of osteogenic medium. Results are representative of 3 experiments performed. (e) BMP2 increases Snail/Slug-YAP/TAZ complex formation. Calvarial osteoblast progenitors isolated from E17.5 mice were treated with 100 ng/ml BMP2 for 2 h. Endogenous Snail/Slug-YAP/TAZ complexes were detected by immunoprecipitation. Results are representative of 3 experiments performed. (f) In situ proximity ligation assay detection of endogenous Snail/YAP/TAZ and Slug/YAP/TAZ interactions in human SSCs. siRNAs: siGFP, siSNAIL, siSLUG or siTEADs(1–4) were transfected, respectively. Nuclei were counterstained with DAPI (blue). The detected interaction sites are marked by fluorescent dots (red). Scale bar: 5μm. Results are representative of 3 experiments performed.

Article Snippet: Human or mouse TEF-1, TEF-3, TEF-4 and TEF-5 (TEAD1–4) siRNAs, and human SNAIL or SLUG siRNAs were obtained from Santa Cruz Technology.

Techniques: Western Blot, Isolation, Transduction, Immunoprecipitation, Transfection, Cell Culture, In Situ, Proximity Ligation Assay