Figure 4.

DOCK8 expression analysis. (a) Western blot analysis of whole PBMC lysates shows DOCK8 expression in patient II.2 and not in patient II.3 with two different DOCK8 antibodies (immunogen indicated in brackets; aa: amino acid); Actin as a loading control. Full-length western blots are provided in the Supplementary Appendix (Supplementary Fig. S4). (b) Flow cytometry of patient II.2 showed DOCK8 expression in majority of NK cells and T cells but no DOCK8 expression in B cells. All cell subsets of patient II.3 lack DOCK8 expression. Gray area: unstained; dashed line: isotype control; solid line: DOCK8 staining. (c) T cell subsets defined by naïve T cells (CCR7⁺CD45RA⁺), central memory T cells (CCR7⁺CD45RA⁻), effector memory T cells (CCR7⁻CD45RA⁻) and T_EMRA cells (CCR7⁻CD45RA⁺) showed no DOCK8 expression in T_EMRA and naïve T cells and DOCK8 expression in majority of central and effector memory T cells of patient II.2 compared to DOCK8 expression in all T cell subsets of a healthy control. (d) Sequencing of cDNA reveals double peaks in chromatograms of T and NK cells of patient II.2 indicating wildtype (black letters) and altered (red letters) transcripts. cDNA chromatogram of B cells shows only single peaks indicating altered transcripts.