Figure 5.
WES of DNA samples and functional assays to identify variations in the JAK-STAT pathway. (A) The inhibitory effect of Rux on p-STAT3 and p-STAT5 in WBCs from healthy donors following stimulation with patient serum. The findings were confirmed in 3 independent experiments using cells from different donors. (B) Filtering method for pHLH, MAS/HLH susceptibility, and JAK-STAT pathway variation and specific variation of the MAS/HLH susceptibility and JAK-STAT pathway in 10 patients. (C-D) Representative immunoblot of STAT5 phosphorylation in transfected 293T cells treated with PBS or PDGF-BB (100 ng/mL) for 15 minutes. (E) Representative immunoblot of STAT3 phosphorylation in transfected 293T cells treated with PBS or IL-7 (100 ng/mL) for 15 minutes. (F) Representative immunoblot of STAT3 phosphorylation in transfected 293T cells treated with PBS or IL-23 (100 ng/mL) for 15 minutes. (G) Representative immunoblot of STAT3 phosphorylation in transfected 293T cells treated with PBS or IL-11 (100 ng/mL) for 15 minutes. (H) Representative immunoblot of STAT3 phosphorylation in transfected 293T cells treated with PBS or IL-10 (100 ng/mL) for 15 or 60 minutes. (I) Representative immunoblot of STAT1 and STAT3 phosphorylation in transfected 293T cells treated with PBS or IL-27 (100 ng/mL) for 15 minutes. (J) Quantitative PCR (qPCR) of CXCL10 in 293T cells transfected with WT or mutant PIAS4 plasmids and stimulated with IFN-γ (50 ng/mL) for 8 hours. (K) qPCR of CXCL11 in 293T cells transfected with WT or mutant SOCS4 plasmids and stimulated with IFN-α (1000 U/mL) for 8 hours. (L) The effect of activation of p-STAT3/5 on WBCs from patients and controls upon stimulation. (M) The effect of upregulation of CXCL10/CXCL11 in WBCs from patients and controls upon stimulation. ∗∗P < .01; ∗∗∗∗P < .0001. HC, healthy control; MFI, median fluorescence intensity; PBS, phosphate-buffered saline; Rux, ruxolitinib.

WES of DNA samples and functional assays to identify variations in the JAK-STAT pathway. (A) The inhibitory effect of Rux on p-STAT3 and p-STAT5 in WBCs from healthy donors following stimulation with patient serum. The findings were confirmed in 3 independent experiments using cells from different donors. (B) Filtering method for pHLH, MAS/HLH susceptibility, and JAK-STAT pathway variation and specific variation of the MAS/HLH susceptibility and JAK-STAT pathway in 10 patients. (C-D) Representative immunoblot of STAT5 phosphorylation in transfected 293T cells treated with PBS or PDGF-BB (100 ng/mL) for 15 minutes. (E) Representative immunoblot of STAT3 phosphorylation in transfected 293T cells treated with PBS or IL-7 (100 ng/mL) for 15 minutes. (F) Representative immunoblot of STAT3 phosphorylation in transfected 293T cells treated with PBS or IL-23 (100 ng/mL) for 15 minutes. (G) Representative immunoblot of STAT3 phosphorylation in transfected 293T cells treated with PBS or IL-11 (100 ng/mL) for 15 minutes. (H) Representative immunoblot of STAT3 phosphorylation in transfected 293T cells treated with PBS or IL-10 (100 ng/mL) for 15 or 60 minutes. (I) Representative immunoblot of STAT1 and STAT3 phosphorylation in transfected 293T cells treated with PBS or IL-27 (100 ng/mL) for 15 minutes. (J) Quantitative PCR (qPCR) of CXCL10 in 293T cells transfected with WT or mutant PIAS4 plasmids and stimulated with IFN-γ (50 ng/mL) for 8 hours. (K) qPCR of CXCL11 in 293T cells transfected with WT or mutant SOCS4 plasmids and stimulated with IFN-α (1000 U/mL) for 8 hours. (L) The effect of activation of p-STAT3/5 on WBCs from patients and controls upon stimulation. (M) The effect of upregulation of CXCL10/CXCL11 in WBCs from patients and controls upon stimulation. ∗∗P < .01; ∗∗∗∗P < .0001. HC, healthy control; MFI, median fluorescence intensity; PBS, phosphate-buffered saline; Rux, ruxolitinib.

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