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Stimulation of human CD4+ve cells with recombinant human IL-2 prior to evaluation of phosphorylated STAT5 by flow cytometry.
a Analysis of JAK3 protein expression in B-cell lines of a healthy control (1), father of the patients (2), patient 1 (3), patient 2 (4) and a ?c-deficient SCID patient (5). Examination of GAPDH protein expression served as a loading control. b Analysis of JAK3 signaling function in B-cell lines of a healthy control, patient 1 (II-1), her father (I-1) and a ?c-deficient SCID patient after stimulation with IL-4 and IL-21. Numbers in the top left and right corners of each histogram in panel B indicate the Mean Fluorescence Intensity (MFI) values of unstimulated and cytokine-stimulated cells, respectively. c Analysis of JAK3 signaling function in CD4+ peripheral blood T-cells of a healthy control, patient 1 (II-1) and patient 2 (II-2) after stimulation with IL-2. Histogram overlays represent intracellular levels of phosphorylated STAT5 in CD4+ T-cells without stimulation or after stimulation with IL-2 or IL-6. Numbers in the left top corner and middle part of each histogram indicate percentages of cells with a positive staining for pSTAT5 following stimulation with IL-2 or IL-6, respectively, while numbers in right corner constitute percentages of pSTAT5-positive unstimulated control cells.
From: Ban SA, Salzer E, Eibl MM, Linder A, Geier CB, Santos-Valente E, Garncarz W, Lion T, Ott R, Seelbach C, Boztug K, Wolf HM. Combined immunodeficiency evolving into predominant CD4+ lymphopenia caused by somatic chimerism in JAK3.
J Clin Immunol. 2014 Nov;34(8):941-53.