Figure 2.

JR-AB2-011 addressed the mTORC2 specific Akt and NDRG1 signaling. Representative Western Blot analysis of total Akt (60 kDa), pAkt^Ser473 (60 kDa), and pNDRG1 (58 kDa) in human melanoma (MelIm) (A) after treatment with 50 µM and 250 µM JR-AB2-011 indicates a dose-dependent reduction in anti-apoptotic Akt signaling in melanoma cells. Representative Western Blot analysis of Caspase 3 (35 kDa) and activated Caspase 3 (cCaspase 3, 17 kDa) indicates no influence of JR-AB2-011 on classical apoptosis in melanoma cells (MelIm) (B). Untreated as well as cytochrome c–treated (Cyt c) Jurkat cells were used as negative and positive controls. Densitometry reveals up to 50% reduction of pAkt^Ser473 phosphorylation as determined by ratio of pAkt^Ser473/Akt (C). Further, phosphorylation of NDRG1 was dose-dependently affected in melanoma cells. Densitometry reveals up to a 62% reduction of NDRG1 phosphorylation (D). 20 μg of protein lysates were separated by SDS-PAGE in three independent experiments, displayed are cropped blots; for full gels see Supplementary Data. PDI was used as a loading control.