Figure 4.

STAT3 and STAT6 in nuclear extracts. PBMC (1–2 × 10⁷) from different probands (wildtype Q551 or variant R551) were stimulated with IL-4 or IL-13. Nuclear extracts were obtained according to the EMSA protocol. Immunoprecipitation was performed using anti-STAT3, followed by western blotting and immunostaining with either p-Tyr or ant-STAT3. Staining with Tyk2 served as a control. In (A) and (B) cells were stimulated for 10 min. In (C) and (D) a time course has been performed. (A). PBMCs 1= WT unstimulated.; 2= WT/ IL-4, 3= WT/ IL-13; 4= R551 unstimulated.; 5= R551/ IL-4; 6= R551/ IL-13; (B). 293 cells 1= unstimulated.; 2= IL-4; 3= IL-13; (C). Time-course experiment IL-4. 1= WT 0 min, 2= WT 1 min, 3= WT 5 min, 4= WT 15 min, 5= WT 20 min, 6= R551 0 min, 7= R551 1 min, 8= R551 5 min, 9= R551 15 min, 10= R551 20 min; (D). Time-course experiment IL-13. 1= WT 0 min, 2= WT 1 min, 3= WT 5 min, 4= WT 15 min, 5= WT 20 min, 6= R551 0 min, 7= R551 1 min, 8= R551 5 min, 9= R551 15 min, 10= R551 20 min. PBMC = Peripheral blood mononuclear cell; WT = wildtype.