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Fig. 3 | Cancer & Metabolism

Fig. 3

From: The PD-L1 metabolic interactome intersects with choline metabolism and inflammation

Fig. 3

Consequences of Chk-α and PD-L1 downregulation on metabolites. a Representative high-resolution 1H MR spectra obtained from the aqueous phase of MDA-MB 231 cells. Spectra are displayed from untreated cells (black), cells transfected with 100 nM luciferase siRNA (light gray), cells transfected with 100 nM Chk-α siRNA (orange), cells transfected with 100 nM PD-L1 siRNA #1 (blue), and cells transfected with a mixture of 50 nM PD-L1 #1 and 50 nM Chk-α siRNA (red). All spectra were plotted on the same vertical scale and acquired with identical experimental parameters. GPC, glycerophosphocholine; PC, phosphocholine; Cho, choline; GSH, glutathione; GSSG, oxidized glutathione; MTA, S-methyl-5′-thioadenosine. b Metabolic heat map, generated from quantitative analysis of high-resolution 1H MR spectral data of the aqueous phase, displaying differences in the metabolic profile of MDA-MB-231 cells. The heat map displays metabolites from untreated cells, cells transfected with 100 nM luciferase siRNA for 48 h, cells transfected with 100 nM Chk-α siRNA for 48 h, cells transfected with 100 nM PD-L1 siRNA #1 for 48 h, and cells transfected with a mixture of 50 nM PD-L1 #1 and 50 nM Chk-α siRNA for 48 h. Heat maps were created using the MATLAB software (MATLAB R2012b, MathWorks) to visualize the metabolic patterns. Due to the high dynamic range of metabolites, we normalized the highest intensity of a metabolite in each of the four groups to 100%. This normalization provides a dynamic range between 0 and 100%, allowing a better presentation of heat maps. The heat map represents the average of 3–6 replicates per group. The integral area under the peak was normalized to the number of cells for each sample. TSP dissolved in D2O was used as a quantitative reference in the spectral analysis. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, compared to the control group (see also Supplementary Table 1)

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