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. 2022 Apr 19;11(9):1377. doi: 10.3390/cells11091377

Figure 2.

Figure 2

miR-324-5p functionally interacts with Nr3c2 (MR) and Elavl1 (HuR) 3′-UTR. The murine Nr3c2 (MR) and Elavl1 (HuR) 3′-UTR were cloned downstream of the pMIR-REPORT luciferase vector. (A,C) HEK 293T cells were transiently transfected, as described in Materials and Methods Section, with pMIR-luciferase plasmid (pMIR-Luc) fused to Nr3c2 (MR) or Elavl1 (HuR) 3′-UTR (40 ng/well of 96-well plates) and incubated with increasing concentrations (10, 50, 100 nM) of negative control Mimics (CTR Mimic) or of 324-5p Mimics. Luciferase activities were measured 24 h after transfection and normalized to β-galactosidase activities. (B,D) HEK 293T cells were transiently transfected with pMIR-Luc fused to Nr3c2 (MR) or Elavl1 (HuR) 3′-UTR (40 ng/well of 96-well plates) and 100 nM of CTR or 324-5p Mimics or with 100 nM of CTR or 324-5p Inhibitors. Luciferase activities were measured 24 h after transfection and normalized to β-galactosidase activities. Data are means ± SEMs (n = 8). NS = not significant, *** p < 0.001, **** p < 0.0001 compared to luciferase activity of pMIR-Luc fused to Nr3c2 (MR) or Elavl1 (HuR) 3′-UTR without Mimics (A,C) or with CTR Mimic or CTR inhibitor (B,D), arbitrarily set at 100%.