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. 2014 Jun 25;56(2):460–471. doi: 10.3109/10428194.2014.924115

Figure 4.

Figure 4.

PrxIII is a direct target of both miR-26a-5p and miR-23b-3p. (A) HEK293 cells were transfected with miRNA inhibitors specific for miR-26a-5p, miR-23b-3p and miR-26a-5p + miR-23b-3p. Equivalent amounts (30 μg) of whole-cell lysates were separated by SDS-PAGE and analyzed by immunoblotting with antibodies specific for the indicated proteins. (B) HEK293 cells were transfected with miRNA mimics specific for miR-26a-5p, miR-23b-3p and miR-26a-5p + miR-23b-3p. Equivalent amounts (30 μg) of whole-cell lysates were separated by SDS-PAGE and analyzed by immunoblotting with antibodies specific for the indicated proteins. (C) Compiled data of PrxIII protein analysis from three independent experiments after transfection of miRNAs is shown. Columns, mean; bars, ± SD; ∗p < 0.05, ∗∗p < 0.01. (D) Compiled data of PrxIII mRNA analysis from three independent experiments after transfection of miRNAs is shown. Columns, mean; bars, ± SD; ∗∗p < 0.01, ∗∗∗p < 0.001. (E) Predicted human PrxIII-encoded mRNA contains a 3′-UTR element that is partially complementary to the indicated miRNAs. Above: predicted target binding sites for miR-26a-5p and miR-23b-3p, respectively. Below: predicted duplex combination between human PrxIII 3′-UTR-wt/mut and miR-26a-5p, PrxIII 3′-UTR-wt/mut and miR-23b-3p. (F) Luciferase activity of pmirGLO3-PrxIII-wt or pmirGLO-PrxIII-mut in HEK293 cells after miR-26a-5p and miR-23b-3p transfections. Data are shown as mean ± SD based on three independent experiments, ∗p < 0.05, ∗∗p < 0.01.