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. Author manuscript; available in PMC: 2014 Aug 1.
Published in final edited form as: Stem Cells. 2013 Aug;31(8):1574–1583. doi: 10.1002/stem.1395

Figure 5. Itch ligase activity is required for increased osteoblast differentiation of Itch−/− cells.

Figure 5

The bone-derived cells from WT mice were cultured in αMEM medium and passaged for 3 times as bone-derived mesenchymal progenitor cells (B-MPCs). (A) Expression profile of surface markers for mesenchymal stem cells in B-MPCs was examined by FACS. MPC surface markers are CD45−/Sca1+/CD105+/CD44+/CD31−/CD11b−/CD117−. (B) The differentiation potential to osteoblasts, adipocytes and chondrocytes were examined by culturing cells in the appropriate inducing media for 2-4 weeks. (C) Bone-MPCs were infected with retroviral supernatant containing GFP, Itch-C830A mutant or WT Itch virus. Cells were cultured in the osteoblast-inducing medium for 5 days and stained for ALP activity. The % of ALP+ area/total plate area was measured. Values are mean ± SD of 3 plates. *P<0.05 vs different groups as indicated.