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. 2014 Feb 24;123(18):2843–2853. doi: 10.1182/blood-2013-05-502435

Figure 2.

Figure 2

Analysis of 2-DG uptake, the expression of GLUT1 and HK, and levels of intracellular G6P, lactate, and ATP in G6PT-deficient neutrophils of GSD-Ib patients. Annexin V–depleted peripheral blood neutrophils isolated from HDs and GSD-Ib patients were used in the study. For quantitative RT-PCR, data represent the mean ± SEM for HDs (n = 10) and GSD-Ib patients (n = 12). (A) Uptake of 2-DG. Data represent the mean ± SEM for HDs (n = 3) and GSD-Ib patients (n = 3). (B) Quantification of mRNA of GLUT1 by real-time RT-PCR. (C) Western blot analysis of protein extracts using antibodies against GLUT1 and HK3. Each lane contains 50 μg of protein. (D) Quantification of GLUT1 and HK3 protein levels by densitometry. Data represent the mean ± SEM for HDs (n = 8) and GSD-Ib patients (n = 7). (E) Confocal analysis of GLUT1 (green fluorescence), pan Cadherin membrane staining (red fluorescence), and DAPI nuclei staining (blue fluorescence) at original magnification ×630. (F) Quantification of HK1, HK2, and HK3 mRNA by real-time RT-PCR. (G) Quantification of G6P, lactate, and ATP. Data represent the mean ± SEM for HDs (n = 13) and GSD-Ib patients (n = 12). **P < .005.