RhoG depletion does not affect lymphocyte activation nor cytokine production. (A) Graph comparing phosphorylated ERK1/2 in T cells upon stimulation with anti-CD3/hICAM1, anti-CD3–coated P815 cells, and PMA/I. Cells were analyzed by flow cytometry after intracellular staining with antibody recognizing phosphorylated ERK1/2 (Thr202/Tyr204). (B) ERK1/2 phosphorylation in WT and RHOG KO NK-92 cells upon stimulation. Analysis was performed similar to panel A. (C-D) Upregulation of CD69 on the surface of NK-92 and T cells upon stimulation with plate-bound Abs (anti-NKp30 or anti-CD3) and hrICAM1. The concentration of the titrated Ab is indicated on the x-axis. (E-F) Graphs show pathologically high levels of IFN-γ (E) and CXCL9 (IFN-γ–inducible chemokine) (F) in the plasma samples of the index patient (Pat) and healthy controls (father, mother, and shipment control, as indicated on the figure legend). Three samples from unrelated FHL3 patients (Pat FHL3) with active disease and corresponding controls (FHL3 Ctrl) were used for comparison. (G) Production of IFN-γ by WT and RHOG KO NK-92 cells upon stimulation with anti-NKp30+hrICAM1, K562 cells, and PMA/I. Cells were analyzed by flow cytometry after intracellular staining for IFN-γ. (H) Release of IFN-γ by WT and RHOG KO NK-92 cells measured 24 hours after stimulation by enzyme-linked immunosorbent assay in the culture medium. Data shown are representative of 2 independent experiments. P values were calculated using a multiple Student t test. *P < .05; **P < .01.