Figure 4: Nf2 is required for osteogenic differentiation of osteosarcomas.
From: Sox2 antagonizes the Hippo pathway to maintain stemness in cancer cells
(a) Nf2 overexpression in mOS-482 cells increases osteogenic differentiation. Western analysis of Nf2 expression mOS-482 cells transduced with a control or Nf2 retrovirus (left panel). Osteosphere assay (middle panel), and osteogenic and adipogenic differentiation (right panel). *P<0.05 by t-test. Error bars represent mean ± s.d. (b) Nf2 depletion in primary osteoblasts reduces osteogenic differentiation. Analysis of Nf2 in primary osteoblasts transduced with scrambled or two independent shNf2 lentivirus- shNf2-A and shNf2-B. Western analysis (left panel). Osteogenic differentiation (right panel). (c) Depletion of Mst1/2 or Lats1/2 in Nf2-overexpressing cells restores sphere formation. Nf2-overexpressing mOS-482 cells were transfected with control (scrambled) or Mst1/2 siRNAs or Lats1/2 siRNAs. Western analysis (upper panel) and sphere assay (lower panel). *P<0.05 by t-test. Error bars represent mean ± s.d. (d) Control or Nf2-overexpressing mOS-482 cells were transduced with myc-tagged YAP (WT), YAP (5SA—constitutively active YAP mutant) or YAP (94SA—YAP mutant deficient in TEAD-binding) retroviruses. Western analysis (upper panel). Osteogenic differentiation (lower panel).
