Fig. 4: TMCC3-induced AKT activation likely does not involve PI3K/PDK1/mTORC1 or mTORC2 pathway. | Oncogene

Fig. 4: TMCC3-induced AKT activation likely does not involve PI3K/PDK1/mTORC1 or mTORC2 pathway.

From: Transmembrane and coiled-coil ___domain family 3 (TMCC3) regulates breast cancer stem cell and AKT activation

Fig. 4

AS-B634 cells were transduced with shRNA-TMCC3 #A, #B, or shRNA-control for 3 days. MCF7 cells were transiently transfected with TMCC3 in the presence (+) or absence (−) of 10 μg/ml human insulin for 5 min after 12 h serum starvation. Levels of phosphorylated AKT (S473 and T308), p85 of PI3K and PDK1, and total protein in TMCC3 silenced AS-B634 (a) and TMCC3 overexpressing MCF7 (b), were determined by western blotting. Relative intensity was quantified and normalized to non-phospho protein or GAPDH. c Subunit p85 phosphorylation and p110-α membrane translocation were determined in TMCC3 overexpressing MCF7 cells. Integrin β1 and GAPDH served as membrane and cytoplasmic markers, respectively. d, e mTORC1 (Raptor and mTOR) and mTORC2 (Rictor and SIN1) activation were assessed in TMCC3 silenced AS-B634 and TMCC3 overexpressing MCF7 cells.

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