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. 2021 Jul 14;24(2):229–244. doi: 10.1093/neuonc/noab175

Fig. 4.

Fig. 4

The effects of NAMPT inhibition on mitochondrial function. (A) Western blot analysis of SirT1 expression in the GSC262, GSC811, and GSC5-22 cell lines. (B) Assessment of the oxygen consumption rate (OCR). An Agilent Seahorse assay was used to determine the effect of KPT-9274 on the basal respiration, maximum respiration, and spare respiratory capacity of the GSC262, GSC811, and GSC5-22 cell lines and the effect of adding 500 µM NMN, a pathway intermediate for the NAD salvage pathway, to reverse the effect of KPT-9274. (C) Immunofluorescence assays for mitochondrial morphologies, numbers, and membrane potentials in GSC262, GSC811, and GSC5-22 cells exposed to KPT-9274 or the vehicle and probed with 4′,6-diamidino-2-phenylindole (DAPI), Tom20-Alexa488, and MitoTracker Red CMXRos. Scale bar = 15 µm (D) Flow cytometric assessments of the mitochondrial potential of the GSC262, GSC811, and GSC5-22 cell lines exposed to 0.1 or 1.0 µM KPT-9274 or the vehicle and measured by staining with 2 µM JC-1 dye.