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. Author manuscript; available in PMC: 2015 Jun 4.
Published in final edited form as: Nat Commun. 2014 Dec 4;5:5641. doi: 10.1038/ncomms6641

Figure 4. Detection of IL-21 mRNA in CD4 T cells.

Figure 4

Lymphocytes isolated from a spleen sample of two subjects suffering from idiopathic thrombocytopenic purpura (ITP) (a, b) and one subject who underwent surgery for traumatic splenic rupture (c). Cells were incubated with culture medium alone or stimulated overnight with PMA/ionomycin, with addition of brefeld in and monensin only for the ICS assay. Following incubation, cells were stained either following a standard ICS protocol or with the new flow-RNA FISH protocol for dead cell dye, surface lineage markers and either cytokine-specific antibodies or cytokine-specific probe sets. (d) Collective data on IL-21 mRNA expression after PMA/Ionomycin stimulation in CD4 T cells from 6 negative control and 2 ITP subjects.