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. 2017 Nov 2;18(11):2305. doi: 10.3390/ijms18112305

Figure 3.

Figure 3

Inhibitory effects of BMS-754807 and NVP-AEW541 on IGF2-induced signaling. (A) IGF1R/IR Inhibitors reduce pAKT. TNBC cell cultures were serum starved for 3 h, then incubated with BMS-754807 or NVP-AEW541 for 1 h. IGF2 (100 ng/mL) was added to the serum-free, and phenol red-free media for 20 min. Total protein was isolated, separated and transferred to PVDF membranes. Detection of Akt (1:1000; Cell Signaling #4685), pAkt (1:1000, Cell Signaling #4060) were evaluated using western immunoblot analysis; (B) IGF1R/IR antagonists slightly reduce pS6 but not pMAPK in TNBC cells. TNBC cultures were serum starved, exposed to IGF1R/IR inhibitors followed by IGF2. The effects of this treatment on MAPK (1:1000, Cell Signaling #9102), pMAPK (1:1000, Cell Signaling #4370), S6 (1:2000, Cell Signaling #2217), and pS6 (1:2000, Cell Signaling #4858) was done as per manufacturer’s recommended protocols (Methods). CON = control vehicle-treated cells. BMS + IGF2 = cells treated with BMS-754807 and IGF2. NVP + IGF2 = cells treated with NVP-AEW541 + IGF2. Western immunoblots are representative of at least three independent experiments.