MicroRNA-19a Targets Fibroblast Growth Factor-Inducible Molecule 14 and Prevents Tubular Damage in Septic AKI

Figure 1

Fn14 is a bona fide target of miR-19a. (a) Predicted miR-19a target sequence in wild-type Fn14-3 UTR (Fn14-WT) among mammalian orthologs and mutant Fn14-3 UTR containing mutated nucleotides in the miR-19a seed-pairing region (Fn14-Mut). (b) RT-qPCR evaluating levels of Fn14 mRNA expression in murine MCT and RAW 264.7 cells transfected with mock (Mock), negative control (NC), and miR-19a mimics (miR-19a) for 48 h, respectively. , one-way ANOVA, post hoc comparisons, and Tukey’s test. (c) Western blotting measuring abundance of Fn14 protein in murine MCT and RAW 264.7 cells transfected with mock (Mock), negative control (NC), and miR-19a mimics (miR-19a) for 48 h, respectively. GAPDH was used as the internal control of cytoplasmic extractions. (d) Luciferase assays of Fn14-3 UTR activity in murine MCT and RAW 264.7 cells cotransfected with WT or Mut Fn14-3 UTR reporter constructs and mock (Mock) and negative control (NC) as well as miR-19a mimics (miR-19a) for 48 h, respectively. and , one-way ANOVA, post hoc comparisons, and Tukey’s test. (e) RT-qPCR assessing abundance of Fn14 mRNA expression in murine MCT cells with mock (Mock) and negative control (NC) as well as miR-19a mimic (miR-19a) transfection in the presence or absence of miR-19a inhibitor (miR-19a in) cotransfection, respectively. , one-way ANOVA, post hoc comparisons, and Tukey’s test. (f) Western blotting analyses comparing levels of Fn14 protein expression in murine MCT cells with mock (Mock) and negative control (NC) as well as miR-19a mimic (miR-19a) transfection in the presence or absence of miR-19a inhibitor (miR-19a in) cotransfection, respectively.

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