Figure 3.

Real-time changes in metabolite peak intensities during 24 h primary CLL NMR time-course. (a) Representative superimposed fragments of spectra of a 24-h time course is shown for some metabolites. The first spectra were colored in red and the last spectra in blue. The black bar indicates the location of each peak that was used for kinetic analysis. Graphs show the intensity difference between the first and the following peaks from the spectra acquired over 24 h for 11 primary CLL samples. (Additional metabolites are shown in Supplementary Figure S7). (b) Schema of an experiment that demonstrates metabolic plasticity of CLL cells. Primary-CLL mononuclear cells were isolated from peripheral blood and incubated for 24 h in normoxia. Then the sample was split into two, one-half was analyzed in the NMR for 24 h (hypoxia) (first cycle) and the other half of the sample was incubated for 24 h in a hypoxic incubator, then for another 24 h in normoxia and finally analyzed in the NMR (hypoxia) for a further 24 h (2nd cycle). (c) Viability data for five primary CLL samples following completion of NMR after having undergone either one or two hypoxic cycles. (d) Representative NMR time-course data for one CLL sample of N=6. Intensity change for lactate, glucose, glutamine and alanine are shown for the cells during the first and the second hypoxic cycle. The dashed line represents oxygen concentration in the NMR tube during the experiment. (Additional metabolites shown in Supplementary Figure S8. Kinetic values corresponding to the time course are shown in Supplementary Table S2).