Fig. 7: TNIK is a CD27 downstream adaptor molecule and regulates Wnt signaling.

a ImageStream X analysis of Numb segregation in mitotic WT and KO p14 T cells after 48 h of co-culture with H8-DCs in the presence of 20 μg ml⁻¹ blocking FR70 Ab or IgG (175–333 mitotic cells). Numb segregation per culture replicate was analyzed, and frequencies of asymmetric division (AD) vs symmetric division (SD) were assessed. Fold change of SD normalized to WT IgG control is depicted. Pooled data from two independent experiments (1–2 replicates per timepoint). b–d In total, 10⁵ Cd45.1⁺ Tnik^WT (WT) or Tnik^−/− (KO) p14 T cells from the spleen were AdTf into naive Cd45.2⁺ mice previous to infection with 10⁴ pfu LCMV-WE. Mice were injected i.p. with 300 μg FR70 mAb or IgG on days −1/+2/+5. b–d Analysis day 6 p.i. (n = 3 mice per group). b FACS-sorted p14 T cells stained for β-catenin. Dotplot shows frequency of nuclear localization analyzed by ImageStream X. c ΔMFI of Eomes and d frequency of KLRG1⁺CD127⁻ p14 SLECs (spleen). e, f Live imaging of dividing αCD3/VAR (n = 183) or αCD3αCD28 (n = 170) in vitro-activated human naive CD8⁺ T cells (T_N) (two donors, two independent experiments): nucleus (NucBlue) and mitochondira (TMRM) stain, merge and brigh field (BF) of representative mitotic cells; scale bar 10 μm. Frequency of AD vs SD of total (n = 170-182) dividing cells is shown. g Histogram showing TCF-1 expression of αCD3/VAR (black lines) or αCD3αCD28 (red lines) activated T_N cells and isotype controls (iso; dashed lines). h Histogram showing CD27 expression (gray solid line), isotype control (gray dotted line), purified CD27⁺ (black line), and CD27⁻ (red line) HIV gag-specific human CD8⁺ T cell clone. i CD27⁺ (black/gray dots) and CD27⁻ (red/light red dots) T cells cultured under 0.3 μg ml⁻¹ OKT3 and 10 μg ml⁻¹ CD27 blocking mAb or IgG. Percentage of cells positive for nuclear TNIK or β-catenin is depicted (three independent experiments; 4–12 replicates; 60–79 cells per replicate); scale bar 10 μm. j TCF/LEF lentiviral reporter assay: luminescence ratio (CD27⁺/CD27⁻) relative to negative control lentiviral particles. h, i Representative results of two independent experiments depicted. Data displayed as means ± SEM. Statistics: a–d, i one-way ANOVA, f, j two-tailed Student's t test, nonsignificant P > 0.05, *P < 0.05, **P < 0.01, ****P < 0.0001. Source data are provided as a Source Data file.