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. Author manuscript; available in PMC: 2013 Jan 1.
Published in final edited form as: J Neurochem. 2011 Nov 24;120(2):334–346. doi: 10.1111/j.1471-4159.2011.07566.x

Fig 3. Effect of the inhibition of PBEF by FK866 on cell viability and NAD+ and ATP levels of cultured neurons.

Fig 3

(A) FK866 reduced neuronal viability under normal and OGD conditions. For measurement of cell viability under normal condition, neurons were treated with indicated concentration of FK866 for 4 h, and cell viability was measured with MTT assay at 24 h starting from FK866 treatment. For measurement of cell viability after OGD, neurons were subject to OGD for 1.5 h in the presence of FK866. FK866 is present for a total of 4 h starting from OGD, and cell viability was measured with MTT assay at 24 h starting from OGD. Data were collected from 4 independent experiments. (B) Effect of FK866 and NAM on NAD+ levels in cortical neurons. Neurons were exposed to different concentrations of FK866 alone or together with 2 mM NAM for 4 h, and NAD+ levels were measured. Data were collected from 4 independent experiments. (C) Effect of FK866 on NAD+ levels after OGD. Neurons were subjected to 1.5 h OGD in the absence or presence of FK866. FK866 was present for a total of 4 h after OGD. NAD+ was measured 6 h after OGD or wash-away of FK866. Data were the mean values from 4 independent experiments. (D) The effect of incubation time of 15 mM NAM on NAD+ levels in OGD. Neurons were subjected to 1.5 h OGD in the presence of 15 mM NAM, and NAM was continuously present for different periods of time indicated in the figure. NAD+ levels were measured 6 h later following wash-away of NAM. Data were the mean value from 5 independent experiments. (E) Effect of NAD+ and NAM on ATP levels in normal and OGD conditions. Neurons were treated with 15 mM NAD+ or 15 mM NAM and were or were not subject to OGD for 1.5 h at the same time. NAD+ and NAM was continuously present for a total time of 24 h and ATP was measured 6 h later after wash-away of NAD+ and NAM. Data were the mean values from 4 independent experiments. *t-test with p<0.05; **t-test with p<0.005 vs the indicated conditions.