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. 2022 May 13;6(6):e722. doi: 10.1097/HS9.0000000000000722

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Copyright © 2022 the Author(s). Published by Wolters Kluwer Health, Inc. on behalf of the European Hematology Association.

This is an open access article distributed under the Creative Commons Attribution License 4.0 (CCBY), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 3. — Increased glucose uptake after anti-IgM stimulation of CLL cells. CLL cells were cultured for 48 hours with either anti-IgM stimulation or control and the residual concentration of glucose ([glucose]^residual) in the media was assessed by colorimetric assay. (A) The [glucose]^residual was lower when CLL cells (n = 26) were stimulated with anti-IgM than when stimulated with isotype control indicating greater glucose uptake with anti-IgM stimulation. (B) The [glucose]^residual was correlated with the percentage of CLL cells that exhibited intracellular calcium flux after IgM stimulation (n = 26). There was a significant correlation between [glucose]^residual and the proportion of CLL cells exhibiting calcium flux, with samples where the majority of cells mobilized calcium also showing the highest degree of glucose uptake. (C) The concentration of lactate ([lactate]) in the media after 48 hours culture was assessed by colorimetric assay. Samples that exhibited calcium flux after anti-IgM stimulation and high glucose uptake (n = 8) had a significant increase in the [lactate] in the media at the end of the culture period compared with nonresponders (n = 6). (D) CLL cells were stimulated by anti-IgM or isotype control for 24 hours and the expression of a panel of glycolytic enzymes was assessed by immunoblot. Representative blots of HK2, GLUT3, PFKM, PFKP, hnRNPA1, PTBP, ENOL-1, and LDHA after anti-IgM stimulation or isotype control. (E) Representative blots comparing the impact of anti-IgM stimulation on HK2 and GLUT3 expression in calcium flux responders and nonresponders. (F) There was no increase in TOMM20 expression in CLL cells stimulated with anti-IgM or isotype control (n = 6). (G) 24 hours anti-IgM treatment resulted in increased respiratory reserve (n = 5). CLL = chronic lymphocytic leukemia; LDHA = lactate dehydrogenase.