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. 2019 Mar 29;14(3):e0214301. doi: 10.1371/journal.pone.0214301

Fig 3. miR-451a inhibits translation of Sparc mRNA.

Fig 3

(A) Global polysome profile of C2C12 cells 48 h after transfection either with Ctrl siRNA or miR-451. (B) The relative distribution of Sparc mRNA (and housekeeping gene Gapdh mRNA) was studied by RT-qPCR analysis of RNA in each of 12 polysome fractions. (C) Top, schematic of the dual-luciferase reporter plasmids psiCHECK2, the control vector expressing renilla luciferase (RL) and the internal control firefly luciferase (FL); the test vector psiCHECK2-Sparc-3′ contains the Sparc 3′UTR downstream of the RL coding region and the psiCHECK2-Sparc-3′-Mut contains the Sparc 3′UTR downstream of the RL coding region with mutated target site for miR-451a. Bottom, C2C12 cells were transfected with either miR-451a or Ctrl siRNA and either psiCHECK2, psiCHECK2-Sparc 3’ or psiCHECK2-Sparc 3’mut; the ratio of RL activity to FL activity was calculated 16 h after transfection. The relative RL/FL ratios of miR-451a-transfected cells relative to RL/FL of Ctrl siRNA-transfected cells are indicated. Data represent the means ± SEM from 3 independent experiments. Significance (P) is indicated.