Fig. 5
Nuclear translocation of NFAT1 is impaired by variants in IP3R3 following TCR but not PMA + ionomycin stimulation. PBMCs were incubated with anti-CD3 and anti-CD28 (both 10 µg/mL) antibodies on ice before cross-linking was induced for 30 min at 37 °C by adding 20 µg/mL of a cross-linking antibody for TCR stimulation (aCD3 + aCD28). PMA + ionomycin (PI) stimulation was performed simultaneously as a positive control. Nuclear translocation was assessed by ImageStream analysis in CD4+ T cells. A Representative images of unstimulated and TCR-stimulated cells from all the individuals. B Histograms of the Similarity Score as a measure of nuclear NFAT1 localization for unstimulated, TCR-stimulated, and PI-stimulated CD4+ T cells. C Percentage of unstimulated, TCR-stimulated, and PI-stimulated CD4+ T cells with nuclear NFAT1. D Median Similarity Score of unstimulated, TCR-stimulated, and PI-stimulated CD4+ T cells. Number of independent experiments: n = 2 with 4 different HDs for unstimulated and TCR-stimulated cells, n = 1 with 2 different HDs for PI-stimulated cells. Individuals were sampled at different time points. The values are presented as the mean + SEM. One-way ANOVA was performed with a multiple comparison test
