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. 2013 Jun 19;8(6):e66789. doi: 10.1371/journal.pone.0066789

Figure 6. Pim-1 is a target of miRNA-15b and protein expression is reduced in Dicer−/− mice.

Figure 6

(A) Western blot analysis of Pim-1 protein from whole tissue homogenates from Dicer+/+ and Dicer−/− mice hearts. (B) Immunohistochemistry showing reduced Pim-1 expression in Dicer−/− cardiac tissue. (C) Western blot analysis of Pim-1 protein from HL-1 cells transfected with si-control, si-Dicer and si-Dicer with miR-15b inhibitor. Quantification of Pim-1 expression was performed using densitometry. (D) Western blot analysis of Pim-1 protein from HL-1 cells transfected with scrambled control and miRNA-15b mimic, (E) Plasmid construct for pLuc-Pim1–3′UTR plasmid. (F) miRNA target reporter luciferase assay with Pim-1 3′UTR. Results were normalized with data obtained from an assay with Renilla luciferase. (G) Luciferase activity in HL-1 cells transfected with a Pim1-mutated 3′UTR and miR-15b mimic.