Extended Data Figure 8: Normal T cell phenotyping results in the SHARPIN-deficient patient ex vivo.

(a) T cell proliferation assay. PMBCs were incubated with Cell Trace Violet, stimulated with anti-CD3/28 or PHA for 72h and analyzed by flow cytometer. (b) Intracellular cytokine staining for Th1, Th2 and Th17 populations. PBMCs were stimulated with PMA (100 ng/ml) and ionomycin (1 μM) for 5h with Brefeldin A. Stimulated cells were surface stained, fixed and permeabilized with BD Cytofix/Cytoperm kit. Cells were further stained for intracellular cytokines and analyzed by flow cytometry. Ctrl: unrelated healthy control, Sister: sister carrying the heterozygous frameshift SHARPIN variant p.Leu74ProfsX86. Representative result of two independent experiments.