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. Author manuscript; available in PMC: 2011 Aug 13.
Published in final edited form as: Mol Cell. 2010 Aug 13;39(3):433–443. doi: 10.1016/j.molcel.2010.07.031

Figure 3. Sin1 is required for mTORC2 dependent Akt hydrophobic motif and Akt turn motif phosphorylation in B cells.

Figure 3

a) The mTORC2 complex integrity in Sin1+/- (WT) or Sin1-/- (KO) Ab-MuLV pre-B cells was determined by immunoprecipitation (IP) of Rictor followed by immunoblotting (IB) of Sin1, Rictor and mTOR.

b) Akt phosphorylation and expression was measured in primary Sin1+/+ or Sin1-/- pro-B cells cultured on OP9 cells with IL-7 and Ab-MuLV transformed Sin1+/- or Sin1-/- pre-B cells. Samples were normalized to total cellular protein and blotted as indicated.

c) FoxO1 phosphorylation and expression was measured in Sin1+/- or Sin1-/- Ab-MuLV pre-B cells. Samples were normalized to total cellular protein and blotted for FoxO1 or FoxO1 p-Thr24.

d) FoxO1 phosphorylation was measured in Sin1+/+ or Sin1-/- pro-B cells cultured with serum and IL-7. The samples were normalized to total FoxO1 protein then blotted for FoxO1 p-Thr24.

e) FoxO1 expression was measured in Sin1+/+ or Sin1-/- pro-B cells. The samples were normalized to total cellular protein.