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

Figure 3

Fn14 contributes to the LPS-induced tubular apoptosis in a miR-19a-dependent manner. (a, b) Western blotting analyses detecting the abundance of Fn14 and CYLD protein in MCT (a) and RAW 264.7 (b) cells with control (Ctrl) siRNA, Fn14 siRNA (si.Fn14), or CYLD siRNA (si.CYLD) transfection, respectively. (c, d) MTT assay measuring cell viabilities of negative control- (NC-) or miR-19a mimic- (miR-19a-) transfected MCT (c) and RAW 264.7 (d) cells exposed to 150 ng/mL LPS (L) for 72 h with control (Ctrl) siRNA, Fn14 siRNA (si.Fn14), or CYLD siRNA (si.CYLD) cotransfection, respectively. , one-way ANOVA, post hoc comparisons, and Tukey’s test. D: DMSO. N.S: not significant. (e, f) DEVDase activity of negative control- (NC-) or miR-19a mimic- (miR-19a-) transfected MCT (e) and RAW 264.7 (f) cells exposed to 150 ng/mL LPS (L) for 72 h with control (Ctrl) siRNA, Fn14 siRNA (si.Fn14), or CYLD siRNA (si.CYLD) cotransfection, respectively. , one-way ANOVA, post hoc comparisons, and Tukey’s test. (g, h) Representative pictures and quantification from TUNEL- and cleaved caspase-3- (c-caspase-3-) staining assay of negative control- (NC-) or miR-19a mimic- (miR-19a-) transfected MCT (g) and RAW 264.7 (h) cells exposed to 150 ng/mL LPS (L) for 72 h with control (Ctrl) siRNA, Fn14 siRNA (si.Fn14), or CYLD siRNA (si.CYLD) cotransfection, respectively. , one-way ANOVA, post hoc comparisons, and Tukey’s test.

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