Figure 5: FA metabolism is critical for activation and rapid proliferation of memory CD4⁺ T cells.

(a) qRT-PCR analyses of the relative expression of the genes encoding fatty acid biosynthesis enzymes in memory Th2 cells at the indicated times after TCR stimulation. The heat map represents the log2 value of the relative mRNA expression level (see colour scale). (b) qRT-PCR analyses of expression of genes encoding the enzymes and transporter in fatty acid uptake and lipolysis programmes in memory Th2 cells collected at the indicated times after TCR stimulation as in a. (c) Representative plots of Bodipy FLC16 in naive CD4⁺ T cells and memory Th2 cells collected at the indicated times after antigenic stimulation are shown. (d) Representative plot of e670 proliferation dye in memory Th2 cells after antigenic stimulation under fatty acid-free conditions. (e–h) Representative plots of Bodipy FLC16 and e670 proliferation dye in stimulated memory Th2 cells in the presence of GW9662 (10 μM) (e,f) or TOFA (10 μM) (g,h). (i) ECAR of memory Th2 cells 48 h after TCR stimulation with or without GW9662 under basal conditions (Time point 0). (**P<0.01, Mann–Whitney U test, n=6 per group). (j) ECAR of memory Th2 cells 48 h after TCR stimulation with or without GW9662 under basal conditions and in response to sequential treatment with Oligomycin, FCCP and Rotenone-Antimycin A. (k) ECAR of memory Th2 cells 48 h after TCR stimulation with or without TOFA under basal conditions. (**P<0.01, Mann–Whitney U test, n=6 per group) (l) ECAR of memory Th2 cells 48 h after TCR stimulation with or without TOFA under basal conditions and in response to sequential treatment with Oligomycin, FCCP and Rotenone-Antimycin A as in j. Three technical replicates were performed for qRT-PCR (a,b). Six technical replicates were performed for Seahorse assay (i–l). The mean values with s.d. are shown (i,k). **P<0.01 Three independent experiments were performed with similar results (a–l).