Figure 3. Increased glycolytic commitment in p53-KO T cells.

Splenic T cells from h3T and h3T-p53 KO mouse were TCR activated for three days and used: A) for determining the fluoresecent glucose (2NBDG) uptake using FACS as detailed in Material and methods. B) to obtain RNA for analyzing the expression of key glycolytic genes (i), HIF1-α, and TIGAR (ii), mitochondrial biogenesis regulator PGC1-α (iii), and pentose phosphate pathway genes (iv). C) for determining basal extracellular acidification rate (ECAR) using seahorse assay bio-analyzer as per manufacturer’s protocol. D) to determine phosphorylation level of S6 protein after intracellular staining and analusis using FACS. (*p<0.05; **p<0.01). Bar diagram on right of each overlay represent cumulative data from different experiments.