CYLD cleavage in primary mouse T cells shows a different kinetic.

<p>(A) CYLD cleavage in primary mouse cells has different kinetics. The activation of primary mouse T cells by CD3 with or without CD28 costimulation leads to the formation of an NH₂-terminal CYLD fragment of approximately 25 kDa. In stark contrast to the rapid kinetics in Jurkat cells, the fragment was first detected after 4 h of stimulation. (B) Primary mouse T cells from <i>PKCθ/β^−/−</i> mice showed normal stimulation-dependent CYLD cleavage, comparable to wild-type control mice. (C) Analysis of the subcellular distribution of a retrovirally introduced CYLD-GFP fusion mutant in unstimulated and stimulated wild-type and <i>PKCθ/β</i>-deficient T cells. (green): the co-localization with Cholera Toxin B stained lipid rafts (red) was monitored using confocal microscopy. Nuclei are stained in blue. Quantification of CYLD-lipid rafts co-localization is shown in the bars in the right panel and reveals a statistically significant decrease in CYLD translocation in unstimulated double knockout cells in comparison to wt control cells. *p<0.05; **p<0.01; ***p<0.001.