Figure S3. Patient Th17 and Th2... | Rockefeller University Press

Figure S3.

$Patient Th17 and Th2 subpopulations and IL-17A and IL-4 production and predicted deleterious (CADD score >20) OSMR variants, including the patient OSMR variant, after deglycosylation treatment. (A) Representative flow plots of CD4+ IL-17A+ T cells from two healthy controls (HCs) (top row) and P1 (bottom row) either untreated (UT; left panels) or treated with PMA and ionomycin for 4 h (right panels). (B) Representative flow plots of the frequency of CD4+ IL-4Ra-positive T cells from two HCs and P1 treated with anti-CD3/CD28 for 5 days. Left dot plot: anti-CD3/CD28-stimulated CD4 (PerCP-conjugated) T cells displayed SSC characteristics consistent with CD4 T cell blasts. Central dot plot: to compensate for autofluorescent CD4 T blast cells, the APC and FITC channel (no antibodies added) were used to define non-autofluorescent CD4 T cells. Right dot plot: from the non-autofluorescent CD4 T cell gate, the fraction of IL-4Ra+ (PE-conjugated) CD4 T cells was defined (IL-4Ra cutoff having been priorly defined as IL-4Ra negativity in the FITC channel along the y axis, not shown). (C) Untreated (UT) and N-deglycosylase PNGase-treated cell lysates of HEK293 cells transfected with empty vector (EV), OSMR WT, and predicted deleterious OSMR variants. The OSMR variant harbored by P1 is marked in red. Source data are available for this figure: SourceData FS3.$

Patient Th17 and Th2 subpopulations and IL-17A and IL-4 production and predicted deleterious (CADD score >20) OSMR variants, including the patient OSMR variant, after deglycosylation treatment. (A) Representative flow plots of CD4⁺ IL-17A⁺ T cells from two healthy controls (HCs) (top row) and P1 (bottom row) either untreated (UT; left panels) or treated with PMA and ionomycin for 4 h (right panels). (B) Representative flow plots of the frequency of CD4⁺ IL-4Ra-positive T cells from two HCs and P1 treated with anti-CD3/CD28 for 5 days. Left dot plot: anti-CD3/CD28-stimulated CD4 (PerCP-conjugated) T cells displayed SSC characteristics consistent with CD4 T cell blasts. Central dot plot: to compensate for autofluorescent CD4 T blast cells, the APC and FITC channel (no antibodies added) were used to define non-autofluorescent CD4 T cells. Right dot plot: from the non-autofluorescent CD4 T cell gate, the fraction of IL-4Ra⁺ (PE-conjugated) CD4 T cells was defined (IL-4Ra cutoff having been priorly defined as IL-4Ra negativity in the FITC channel along the y axis, not shown). (C) Untreated (UT) and N-deglycosylase PNGase-treated cell lysates of HEK293 cells transfected with empty vector (EV), OSMR WT, and predicted deleterious OSMR variants. The OSMR variant harbored by P1 is marked in red. Source data are available for this figure: SourceData FS3.