Fig. 3: Proteomic analysis of resting and activated CD4⁺ T cells from CCT8T^+/+ and CCT8T^−/− mice.

a T cells activated in vitro by CD3/CD28 cross-linking. Expansion index and normalized cell viability. b Expression of individual CCT subunits, and c the CCT substrates actin and tubulin measured by mass spectrometry. d Nuclear actin filaments in activated CD4⁺ T cells in the presence of absence of CK-666, an inhibitor of Arp2/3. STED images with segmentation of the nuclear actin filaments (right panels), and e frequency of nuclear actin filaments detected in CCT8^T+/+ (white bars) and CCT8^T−/− T cells (grey bars). The data display an average of 70 nuclei inspected in each of three independent experiments. f Differentially expressed proteins in resting (0 h) and activated (24 h) T cells. GO analysis of cellular components enriched in CCT8^T+/+ CD4⁺ cells at 0 h (g) and 24 h (h) after stimulation. i The normalized MFI (MFI-positive population/MFI-negative population) of total STAT1 (tSTAT1) and phosohorylated STAT1 (pSTAT1) at 0 h and at 24 h upon activation in vitro by CD3/CD28 cross-linking. ^**p < 0.01, ^***p < 0.001, ^****p < 0.0001. Data were calculated by Student’s t-test (a, e and i) correcting for multiple comparisons (Holm-Sidak method; (a)) and ANOVA with post hoc test (b, c and f). Data shown in bar graphs represent mean ± SD values of a single experiment representative of two independent experiments with three replicates each, and results in heat maps and GO analyses are from two independent experiments with three replicates each. See also Fig. S2.