1. DGKζ and Cbl-b deficient T Cells have enhanced ERK1/2 and IκBα phosphorylation.
DGKζ and Cbl-b deficient T Cells have enhanced ERK1/2 and IκBα phosphorylation. A total of 4 × 105 Stemcell Technologies–purified WT, DGKζ−/−, Cbl-b−/−, or DKO naive CD8+ T cells were incubated with biotinylated α-CD3 (5 μg/ml) and α-CD28 (5 μg/ml) and cross-linked with streptavidin (25 μg/ml) for the indicated times. Lysates were immunoblotted for protein levels of (A) phosphorylated ERK (pERK), total ERK (tERK), and β-actin or (B) phosphorylated IκBα (p-IκBα), total IκBα (IκBα), and β-actin. Relative band intensities are indicated below each lane. Representative blots are depicted from one of three independent iterations and ImageJ quantification of the three iterations are displayed ± SEM in the graphs. The β-actin control blot from (A) and (B) is an identical image because the representative images from (A) and (B) were obtained from a single membrane probed with the indicated Abs. *p < 0.05, **p < ns, not significant.
Erin M. Wesley et al. ImmunoHorizons 2018;2:
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