FIGURE 3.

The kinase activity of PKCδ is dispensable for its inhibitory role in TCR-induced activation of NF-κB. A, PKCδ-knockdown Jurkat T cells were transfected with MIGR-GFP-PKCδ-WT (wild type) or MIGR-GFP-PKCδ-KD (kinase-dead) mutant by retroviral transduction. Seventy-two hours later, reconstituted cells were sorted by FACS. The efficiency of reconstitution was examined by immunoblotting with an antibody to PKCδ or β-actin. B and C, PKCδ-knockdown Jurkat T cells, reconstituted Jurkat T cells with PKCδ WT or KD mutants, and control cell lines were stimulated by CD3/CD28 crosslinking. The culture medium was collected and subjected to ELISA for measuring the production of IL-2 (B). The cell lysates were applied to immunoblots with indicated antibodies (C).