Figure 1.
Autosomal dominat STAT1 deficiency in a girl with coccidiodomycosis. (A) Erythematous and verrucous plaque in the right deltoid region at 16 mo of age. (B) Dermal vasculitis of the sole associated with SARS-CoV-2 infection. (C) Inflammatory tumors (arrow) in the soft tissues of the right frontotemporal region. (D) Lytic lesions in the frontotemporal (arrow) bones overlying the soft tissue tumors revealed using CT. (E) Cranial CT shows lytic lesions in the temporal bone overlying the soft tissue tumors (sagittal). (F) Thorax CT shows lytic lesions in the L1, L2, L4, and S1 vertebrae. (G) Thorax CT indicates a lung nodule in the right basal lobe. (H) Thorax CT shows miliary pattern images. (I) Microscopic observation of spherule images after periodic acid–Schiff staining indicated Coccidioides spp. in the soft tissue specimens of the frontotemporal region. Evaluation of the variant of STAT1 K410E (J–M). (J) U3C cells were transfected with plasmids carrying empty vector (mock), WT, K410E, Y701C (a known LOF), or R274Q (a known gain-of-function) mutants at 2 ng/well, along with the reporter plasmids (Cignal GAS Reporter Assay Kit, QIAGEN). The total amount of the STAT1 plasmid was kept constant at 5 ng/well by supplementing with the mock vector. 24 h after transfection, the transfectants were stimulated with IFN-γ at 10 or 1,000 IU/ml for 16 h. Subsequently, the luciferase assays were performed with the Dual-Glo Luciferase Assay System (Promega). The results shown in the bar graphs represent the mean ± SEM from three independent experiments. (K and L) The evaluation of STAT1 variants under conditions where varying amounts of WT (1 or 2 ng) and/or mutant (1, 2, or 3 ng) STAT1 plasmids were cotransfected, with the total plasmid amount maintained at 5 ng/well using the mock vector. The expression of introduced STAT1 was evaluated by immunoblot. GAS reporter activity was measured after stimulation with 1,000 IU/ml IFN-γ. These experiments were performed to assess the effect of STAT1 mutants on WT-mediated GAS induction. The results shown in the bar graphs represent the mean ± SEM from three independent experiments. (M) U3C cells transiently expressing WT or mutant STAT1 were stimulated with 1,000 IU/ml of IFN-γ for 15 min and then subjected to immunoblot analysis. The following primary antibodies were used: anti-STAT1, anti-phosphorylated STAT1 at Tyr-701 (pSTAT1), and anti-β-actin. SARS-CoV-2, severe acute respiratory syndrome coronavirus 2; SEM, standard error of the mean. RLU: relative luminescence units. Source data are available for this figure: SourceData F1.

Autosomal dominat STAT1 deficiency in a girl with coccidiodomycosis. (A) Erythematous and verrucous plaque in the right deltoid region at 16 mo of age. (B) Dermal vasculitis of the sole associated with SARS-CoV-2 infection. (C) Inflammatory tumors (arrow) in the soft tissues of the right frontotemporal region. (D) Lytic lesions in the frontotemporal (arrow) bones overlying the soft tissue tumors revealed using CT. (E) Cranial CT shows lytic lesions in the temporal bone overlying the soft tissue tumors (sagittal). (F) Thorax CT shows lytic lesions in the L1, L2, L4, and S1 vertebrae. (G) Thorax CT indicates a lung nodule in the right basal lobe. (H) Thorax CT shows miliary pattern images. (I) Microscopic observation of spherule images after periodic acid–Schiff staining indicated Coccidioides spp. in the soft tissue specimens of the frontotemporal region. Evaluation of the variant of STAT1 K410E (J–M). (J) U3C cells were transfected with plasmids carrying empty vector (mock), WT, K410E, Y701C (a known LOF), or R274Q (a known gain-of-function) mutants at 2 ng/well, along with the reporter plasmids (Cignal GAS Reporter Assay Kit, QIAGEN). The total amount of the STAT1 plasmid was kept constant at 5 ng/well by supplementing with the mock vector. 24 h after transfection, the transfectants were stimulated with IFN-γ at 10 or 1,000 IU/ml for 16 h. Subsequently, the luciferase assays were performed with the Dual-Glo Luciferase Assay System (Promega). The results shown in the bar graphs represent the mean ± SEM from three independent experiments. (K and L) The evaluation of STAT1 variants under conditions where varying amounts of WT (1 or 2 ng) and/or mutant (1, 2, or 3 ng) STAT1 plasmids were cotransfected, with the total plasmid amount maintained at 5 ng/well using the mock vector. The expression of introduced STAT1 was evaluated by immunoblot. GAS reporter activity was measured after stimulation with 1,000 IU/ml IFN-γ. These experiments were performed to assess the effect of STAT1 mutants on WT-mediated GAS induction. The results shown in the bar graphs represent the mean ± SEM from three independent experiments. (M) U3C cells transiently expressing WT or mutant STAT1 were stimulated with 1,000 IU/ml of IFN-γ for 15 min and then subjected to immunoblot analysis. The following primary antibodies were used: anti-STAT1, anti-phosphorylated STAT1 at Tyr-701 (pSTAT1), and anti-β-actin. SARS-CoV-2, severe acute respiratory syndrome coronavirus 2; SEM, standard error of the mean. RLU: relative luminescence units. Source data are available for this figure: SourceData F1.

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