Fig. 2

PKCθ-E2 mutation impairs PKCθ expression at the protein but not at the mRNA level. a PKCθ mRNA expression of MACS-sorted CD4⁺ T cells isolated from wt or E2^mut mice was analyzed by qRT-PCR. The house keeping gene gapdh was used for normalization. Each symbol represents values obtained of an individual mouse (n = 9), the connecting lines show indicate which samples were analyzed together in one experiment. Statistical significance was determined with paired t-test. b PKCθ protein expression of MACS-sorted CD4⁺ T cells from wild type (wt), PKCθ-deficient (ko) and mice expressing a PKCθ version with mutated exon 2 (E2^mut) was analyzed by SDS-PAGE and immunoblot. Fyn was used as loading control. Representative results of one experiment (of total n = 8) are shown. c Representative western blots of PKCθ and actin expression levels of CD4⁺ T cells isolated from PKCθ-E2^wt and PKCθ-E2^mut mice untreated and after cycloheximide treatment for the indicated time points. The graph summarizes the results of two independent experiments with total of four experimental groups per genotype (mean ± SEM)