Figure 3.

(A) DPP activates AKT, mTOR and NFκB signaling pathways in T4-4 cells. T4-4 cells were cultured on DPP coated non tissue culture plates for 4,8,16, and 24 h. Cells were lysed and equal amounts of proteins were separated by SDS-PAGE, transferred to PVDF membrane, and immunoblotted with antibodies against AKT, p-AKT (Ser 473), p-AKT (Thr 308), p-PDK1 (ser 241), and Tubulin (A). (B) Inhibitor LY294002 causes in-activation of AKT in DPP mediated adhered T4-4 cells. T4-4 cells were pre-treated with LY294002 10 and 50 μm for 30 min and then seeded on DPP substrate for 16 h. After 16 h, cells were lysed and equal amounts of proteins were separated by SDS-PAGE, transferred to PVDF membrane, and immunoblotted with antibodies against p-AKT, total AKT and tubulin. (C) DPP mediated activation of activation of NFκB and mTOR pathways. Total proteins were isolated from the T4-4 adhered cells, lysed and equal amounts of proteins were separated by SDS-PAGE, transferred to PVDF membrane, and immunoblotted with antibodies against p-NFκB, p-mTOR, total NFκB and total mTOR antibodies. Equal loading of the proteins were confirmed by stripping and reprobing the blot with tubulin antibody.