Fig. 3

Role of PKD3 for T cell activation in vitro. a–d CD4⁺ T cells isolated from spleens of either wt or PKD3-deficient mice were stimulated ex vivo with anti-CD3/CD28 antibodies for the indicated time points. IL-2 and IFN-γ expression was assessed either on mRNA level (after 2.5 h, 18 h and 44 h) by qRT-PCR (a, c) or after 44 h in the culture supernatant by luminex methodology (b, d). The house keeping gene gapdh was used for normalization of the mRNA data, and the results are depicted relative to unstimulated cells ex vivo. Secreted IL-2 and IFN-γ are shown relative to wt, with the mean wt value of each experiment set to 100. e, f IL-2 and IFN-γ mRNA expression in naïve sorted CD44⁻CD4⁺ T cells of both genotypes was analyzed by qRT-PCR after sort (unstimulated) and upon in vitro differentiation under non-polarizing (Th0) or Th1-polarizing conditions at the depicted time points. The mRNA data are shown relative to GAPDH. Data are represented as individual values plus mean from 3 independent experiments (n ≥ 8 mice) for a-d and 5 independent experiments (n ≥ 6 mice) for e/f *p < 0.05, ***p < 0.001; two-tailed unpaired t-test followed by Bonferroni’s correction for multiple tests within one graph