Skip to main content
. 2015 Mar 4;8(6):1201–1211. doi: 10.1038/mi.2015.11

Figure 1.

Figure 1

Expression of SMAR1 is regulated by GATA-3. Naive CD4+ T cells were isolated from C57BL/6 mice and activated under various polarizing conditions with plate-bound CD3 and CD28 antibodies. (a) Quantitative real-time mRNA expression of SMAR1 during type 1 helper T cell (Th1), Th2, and Th17 differentiation. (b) Confocal staining for SMAR1 (green) and GATA3 (red) in CD4+ T cells isolated from mice and cultured under Th1 and Th2 differentiation conditions. Relative mRNA expression of SMAR1 was quantified during the course of CD4+ T-cell differentiation to (c) Th2, (d) Th1, and (e) Th17 using real-time PCR. (f) Schematic diagram of the SMAR1 promoter and the regions (not to scale) amplified for the chromatin immunoprecipitation (ChIP) assay. SPr-I and -II correspond to the GATA-binding consensus motifs on SMAR1 promoter. (g) SMAR1 promoter luciferase assay performed by transient transfection of SMAR1 promoter luciferase construct (SPr-I Luc PGL-3 vector) along with GATA-3 expression plasmid in HEK 293 cells. Data represent relative fold change in luciferase activity. (h, i) ChIP assays for GATA-3 and CBP/p300 binding to the SMAR1 promoter SPr-I in CD4+ T cells in naive (N), Th1-, or Th2-polarizing conditions. *P<0.01, **P<0.005. Data are representative of three independent experiments.