Figure 8. A Snail/Slug-YAP/TAZ Axis Regulates SSC Function.

(a) CFU-F generation from Snail^f/f/Slug^+/+/GFP or Snail^f/f/Slug^−/−/Cre SSCs that were transduced with lentiviral GFP, YAP or TAZ constructs. Results are representative of 5 experiments performed.

(b) CFU-F colony counts from (a) (mean ± s.d., n=5 independent experiments). **p<0.01; one-way ANOVA.

(c) Alizarin Red S staining of Snail^f/f/Slug^−/−/Cre or Snail^f/f/Slug^+/+/GFP SSCs transduced with lentiviral GFP, YAP or TAZ, and cultured under osteogenic conditions for 14 d. Results are representative of 3 experiments performed.

(d) Relative mRNA expression of osteogenic markers (Runx2, Osterix, Alp or Bglap2) in cultures from (c) (mean ± s.d., n=3 independent experiments). **p<0.01; one-way ANOVA.

(e) Immunoblotting of YAP/TAZ, Snail/Slug and mutant Snail (N150). C3H10T1/2 cells were transfected with mutant Snail (N150) or empty vector (EV). Results are representative of 3 experiments performed.

(f) Mutant Snail (N150) inhibits proliferation of C3H10T1/2 cells. Proliferative response in cells from (e) were assayed by XTT assay (mean ± s.d., n=3 independent experiments). **p<0.01; unpaired t-test.

(g) Relative mRNA expression of YAP/TAZ/TEAD targets was assessed in cells from (e) (mean ± s.d., n=3 independent experiments). **p<0.01; unpaired t-test.

(h) Mutant Snail (N150) inhibits osteogenesis of C3H10T1/2 cells. Cells from (e) were cultured under osteogenic conditions for 7 d. Osteogenesis was monitored by ALP staining.

(i) Relative mRNA expression of osteogenic markers (Runx2, Osterix, Alp and Bglap2) was assessed in cells from (h) (mean ± s.d., n=3 independent experiments). **p<0.01; unpaired t-test.

(j) Schematic model of Snail/Slug in regulating SSC proliferation and osteogenic differentiation.