Figure 4
miR-720 Represses Cell Cycle-Related Genes.
(a) Relative expression levels of AP-1 family genes in miR-720-overexpressing primary human CD8+ T cells. Data are representative of four independent experiments (n = 4 and 4). (b) Predicted miR-720 targeting sites in the FOSB 3′-UTR. *indicates the seed-sequence mutations. (c) Expression levels of miR-720 in stable miR-720-overexpressing 3T3 cell lines. (d) Luciferase activity in cells with miR-720 seed-sequence mutations at the FOSB 3′-UTR. Data are representative of three independent experiments. (e) FOSB mRNA expression in FOSB siRNA-silenced human primary CD8+ T cells as determined by qPCR. (f and g) Proliferation of human CD8+ T cells after siRNA-mediated Fosb silencing. Data are representative of four independent experiments (n = 4 and 4). (h) Predicted miR-720 targeting sites in the c-Myc 5′-UTR and CDS. * indicates seed-sequence mutations. (i) Myc mRNA expression in miR-720-overexpressing primary human CD8+ T cells compared to control siRNA-transfected cells. Data are representative of three independent experiments (n = 3 and 3). (j) Luciferase activity in cells with miR-720 seed-sequence mutations at the c-Myc 5′-UTR and CDS. Data are representative of three independent experiments (k) c-Myc mRNA levels after siRNA silencing in primary CD8+ T cells as measured by qPCR. (l and m) Proliferation of primary human CD8+ T cells after siRNA-mediated Myc silencing. Data are representative of four independent experiments (n = 4 and 4). Data are represented as mean ± SD in (a, c, d, g, i, j and m). Two-tailed paired (a, g, i and m) and unpaired (d and j) Student’s t-tests were used to determine significance. *p < 0.05; **P < 0.01; ***p < 0.001.
