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. 2013 Sep 23;210(10):2119–2134. doi: 10.1084/jem.20130252

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© 2013 Yang et al.

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Figure 5. — Overexpression of PFKFB3 in RA T cells restores glucose metabolism and protects from apoptosis. CD4⁺CD45RA⁻ T cells were isolated from RA patients, stimulated with anti-CD3/CD28 beads, and 48 h later transfected with PFKFB3 overexpression plasmids (pIRES-PFKFB3-GPF) or control plasmids. Cells were kept in culture for an additional 48 h. Flow cytometric analysis of a representative experiment analyzing GFP expression is shown (A). The following parameters were determined in transfected T cells: (B) PFKFB3 mRNA level by qPCR; (C) lactate production; and (D) intracellular ATP concentration quantified as in Fig. 1. Data from five independent experiments are presented as mean ± SEM. Apoptotic susceptibility was tested in PFKFB3 and control transfected T cells by measuring 7AAD⁺ and Annexin V⁺ cells (E). *, P < 0.05; **, P < 0.01.