miR-34a enhances T cell activation through repressing DGKζ expression.

<p>(<b>A</b>) Schematic representation of CMV promoter-driven miR-34 sponge (SPN-34). (<b>B</b>) Stable Jurkat T cells transduced with miR-34a or SPN-34 were lysed, and DGKζ protein levels were analyzed by a western blot and quantified with a densitometer. (<b>C</b>) DGKα mRNA expression were determined by real-time qPCR (a.u., arbitrary unit; *<i>P</i> < 0.05). (<b>D</b>) Jurkat T cells transduced with miR-34a or SPN-34 were stimulated with C305 (1:50000) overnight, followed by FACS analysis of CD69 expression. Bar graphs display mean ± SEM from three experiments (gMFI, geometric mean fluorescence intensity; unstim, unstimulated; stim, stimulated; ***<i>P</i> < 0.001). (<b>E</b>) Cells were stimulated with C305 (1:10000, 10 min), fixed, and permeablized. Erk1/2 phosphorylation was determined by intracellular staining and FACS analysis. gMFIs were calculated from triplicates of three experiments (mean ± SEM). (<b>F</b>) IL-2 mRNA levels in the indicated cells following 8 hr C305 stimulation were analyzed by real-time qPCR.