Skip to main content
. Author manuscript; available in PMC: 2012 Feb 3.
Published in final edited form as: Immunity. 2009 Dec 18;31(6):941–952. doi: 10.1016/j.immuni.2009.10.008

Figure 1. Characterization of an IL-21 Response Element 3′ of the Prdm1 Gene.

Figure 1

(A) IL-21 induces Prdm1 gene expression. Prdm1 mRNA (means ± SEM of ≥3 independent experiments, total combined samples ≥4) was determined by quantitative RT-PCR 6 hr after treatment with 50 ng/ml IL-21.

(B) Cells were activated with 1 µg/ml anti-CD40 and/or 3 µg/ml anti-IgM-Fab for 3 days, rested 24 hr, and treated with 100 ng/ml IL-21. Prdm1 mRNA (means ± SEM of two independent experiments, total combined samples = 4) was determined 24 hr later by quantitative RT-PCR.

(C) Cells were activated with 1 µg/ml anti-CD40 for 3 days, rested for 24 hr, and treated with 100 ng/ml IL-21 for 24 hr and immunoblotted with antibodies to BLIMP1 or actin. Representative results of three independent experiments.

(D) Schematic of the Prdm1 gene (defined by NM_007548 version 2; version 3 of May 2008 starts 61 bp downstream and lacks the 79 bp second exon) and locations of luciferase reporter constructs. pGL4-pro, minimal promoter (−972 to +183) reporter; K, KpnI; X, XhoI; S, SmaI; open triangle is the luciferase insertion site in exon 1 for R1, R8, R9, and R10.

(E) Luciferase assays in NFS201 cells were performed 18 hr after treatment with 50 ng/ml IL-21.

(F) Shown is luciferase activity 6 hr after treatment with 50 ng/ml IL-21 for NFS201 (means ± SEM of ≥4 independent experiments, total combined samples ≥6) and with 100 ng/ml IL-21 for splenic B cells (means ± SEM of ≥2 independent experiments, total combined samples ≥4).

(G and H) More detailed mapping of the Prdm1 IL-21 response element in preactivated splenic B cells. Shown is luciferase activity 6 hr after treatment with 100 ng/ml IL-21 (means ± SEM of ≥3 independent experiments, total combined samples ≥6).