1. A
4h of treatment
72h of treatment
1mM PI3Kbi - + - - - + + + + + - + - - - + + + + +
untreated
1mM PI3Kai - - + - - + - - + - - - + - - + - - + -
1mM IGF1Ri - - - + - - + - - + - - - + - - + - - +
0.1mM MEKi - - - - + - - + + + - - - - + - - + + +
a-pAkt S473
a-pAkt T308
a-Akt
a-pPRAS40 T246
a-pNDRG1 T346
a-pERK1/2 T202/Y204
a-ERK1/2
a-pS6 S235/236
a-pS6 S240/244
a-S6
a-tubulin 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 B
2h of treatment
72h of treatment
1mM PI3Kbi - + - - - + + + + + - + - - - + + + + +
untreated
1mM PI3Kai - - + - - + - - + - - - + - - + - - + -
0.01mM IGF1Ri - - - + - - + - - + - - - + - - + - - +
0.1mM MEKi - - - - + - - + + + - - - - + - - + + +
a-pAkt S473
a-pAkt T308
a-Akt
a-pPRAS40 T246
a-pNDRG1 T346
a-pERK1/2 T202/Y204
a-ERK1/2
a-pS6 S235/236
a-pS6 S240/244
a-S6
a-tubulin 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21
Suppl. Figure 6: Concomitant inhibition of PI3Kb, IGF1R and MAPK signaling are leading to full long-term pathway blockade.
A) +B) Effects of treatment with the indicated inhibitors as single-agents or in combination on WM (A) or RVH-421 (B) were evaluated by immunoblotting using phospho-specific or total target protein antibodies.
PI3Kbi=rac-KIN-193, PI3Kai=BYL719, IGF1Ri (A)=AEW541, IGF1Ri (B)=Figitumumab-like antibody, MEKi=MEK162