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. Author manuscript; available in PMC: 2017 Sep 1.
Published in final edited form as: Mol Cancer Ther. 2016 Jul 7;15(9):2119–2129. doi: 10.1158/1535-7163.MCT-16-0197

Figure 1.

Figure 1

KPT-9274 inhibits NAMPT and PAK4 and associated signaling pathways in RCC cells.

A. Molecular structure of KPT-9274

B. Cell-free assay of NAMPT activity as a function of KPT-9274 concentration

C. CRISPR-Cas9 splicing out of PAK4 in U-2 osteosarcoma cells showing the expected immunoblot and a shift to the right in the survival curve assessed by MTT assay.

D. NAD+NADH assay: 300 cells/well were plated in 96 well plates (n=4) and incubated for 48h with DMSO or KPT-9274. Assays of total NAD+NADH were performed in RCC cell lines as well as a normal primary renal proximal tubular epithelial cell (RPTEC) line as described in Materials and Methods. Error bars are SD.

*p<0.05 compared to DMSO treated controls. The black sold line indicates treatments significantly different compared to DMSO alone.

E. Immunoblotting of the cell lysates with the indicated antibodies was performed. Cells were plated in 6-well (1000 cells/well) and incubated with different concentrations of KPT-9274 for 72h before they lysed for protein extraction.

The experiments shown are representative of at least three independent repeats.

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