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. 2012 Dec 5;41(2):1273–1283. doi: 10.1093/nar/gks1228

Figure 3.

Figure 3.

Experimental confirmation of interaction between selected miRNAs and their putative 3′-UTR targets and evidence for apparent synergistic effects of miR-195 and miR-382. HeLa cells were transfected with luciferase reporter constructs containing the following 3′-UTR segments: ATP-sensitive inward rectifier K+ channel (Romk2), Na+/K+/2Cl− cotransporter (Nkcc2 or Slc12a1) or Uromodulin or Tamm–Horsfall glycoprotein (Umod or THP). Cells were also transfected with 100 nM of the following oligonucleotides: pre-miR-16 (pre-16), pre-miR-195 (pre-195) and pre-miR-382 (pre-382) or control oligonucleotides (pre-neg). (A) n = 5–10, *P < 0.05 versus control. (B) n = 5–10, *P < 0.05 versus control. (C) n = 13–14, *P < 0.05 versus control. (D) HeLa cells were transfected with luciferase reporter constructs containing the 3′-UTR of Romk2. Cells were also transfected with the following miRNA mimics: pre-195 (30 nM), pre-382 (30 nM), pre-195 (15 nM) + pre-382 (15 nM) or pre-neg control (30 nM) oligonucleotides. In all treatment conditions, the total concentration of miRNA mimics was equal (30 nM). The extent of luciferase suppression was greater with simultaneous treatment with miR-195 and miR-382 mimics, compared with the sum of the effect of each miRNA applied individually. n = 5, *P < 0.05 versus the other groups.