FIGURE 5.

Direct targeting of CD3d, ITK, NFATc3, IL‐2RA, and FOXO1 by overexpression of miR‐182, and polarization of these cells toward FOXP3+ cells (Tregs). A, Comparison between cotransfection of constructs that contain Renilla luciferase reporter gene and 3′UTR of each CD3d, ITK, NFATc3, IL‐2RA, and FOXO1 sequence accompanied by miR‐182–overexpressing vector in HEK‐293t cells. Moreover, miR‐182–overexpressing vector with mutated controls of each gene were used to strengthen the specificity of miR‐182 in targeting of the mentioned targets. The other control group was a mock control which PEZ‐LV105a cotransfected with constructs that contain Renilla luciferase reporter gene and 3′UTR of the mentioned gene. The last control group was psiCHECK‐2 vector that contained Renilla luciferase transfected in HEK293t cells. Luciferase assay represents that miR‐182 is capable of interacting with 3′UTR of CD3d, ITK, NFATc3, IL‐2RA, and FOXO1 sequences cloned downstream of the luciferase ORF and declines luciferase ratios, in contrast to 3′UTR of mutated controls, mock, and FOXP3 (negative control). B, The binding site of CID region of miR‐182‐3p on 3'UTR sequences of CD3d, ITK, NFATc3, and IL‐2RA