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. 2016 Mar 27;7(19):27313–27324. doi: 10.18632/oncotarget.8430

Figure 5. The human 3′UTR of GRβ is regulated by miR144.

Figure 5

The pMirTarget vector containing the 3′UTR of human GRβ was cloned into a luciferase reporter gene (3′UTR GRβ-Luc) (A). The T24 and UMUC-3 bladder cancer cells were transfected with the 3′UTR GRβ-Luc expression construct with mutation in the miRNA binding site for miR181, miR144, or miR33a and was measured by a luciferase assay, and normalized to renilla (B). ***p < 0.001; ****p < 0.0001 (versus WT) (±S.E.; n = 6). The miRNA expression in the UMUC3 and T24 cells was measured using Real-Time PCR (C). *p < 0.05; **p < 0.01 (versus UMUC3) (±S.E.; n = 3). Total RNA was harvested at the time of wounding and 3 hours after from the T24 cells in media containing 10% dialyzed FBS to determine the expression during migration assay for miRNA expression (D). *p < 0.05 (versus T0) (±S.E.; n = 3), and for mRNA expression of GRβ and GRα expression (E). ***p < 0.001 (versus T0) (±S.E.; n = 3). A plasmid containing the human miR144 in the pCMV-MIR vector was transfected in the T24 cells to show how miR144 overexpression affected the expression of GRβ and GRα as measured by Real-time PCR (F). *p < 0.05 (versus T24 Vector) (±S.E.; n = 3).