Figure 3.

MiR-10b directly binds to the 3′UTR of GATA3 and regulates the differentiation of CD4⁺ T cells

(A and C) Schematic representation of the binding site for miR-10b in the GATA3 3′ UTR. (B and D) Luciferase activity in HEK-293T cells co-transfected with the pmirGLO-GATA3-3′ UTR or the pmirGLO-PTEN-3′ UTR sequence, and miR-10b mimic. The results represent the mean ± SD from three independent experiments. Student’s t test; ∗p < 0.05. (E and F) Relative protein expression levels of GATA3 and PTEN in Jurkat cells and U87MG cells, respectively, after transfection with the miR-10b mimic or negative control. The results are presented as the mean ± SD from six independent experiment, ∗p < 0.05. (G and H) The representative expression of GATA3 and PTEN in synovium from RA patients and healthy donors using immunohistochemical staining. (I and K) The percentage of Th1 and Th2 cells in FACS-sorted CD4⁺ T cells from PBMCs of healthy donors (30 donors) and mouse spleens (4 mice) after transfection with human GATA3 siRNA and mouse GATA3 siRNA were analyzed by flow cytometry. (J and L) The percentage of Th17 and Treg cells in FACS-sorted CD4⁺ T cells from PBMCs of healthy donors (30 donors) and mouse spleens (4 mice) after transfection with human PTEN siRNA and mouse PTEN siRNA were analyzed by flow cytometry. CD4⁺ T cells were treated with PMA, ionomycin, and BFA before flow cytometry analysis. Figures are presented as mean ± SD from at least three independent experiments. Student’s t test; ∗p < 0.05, ∗∗p < 0.01.