Figure 6: In vivo siRNA mediated FECH knockdown hastens the development of vemurafenib nephrotoxicity.

Age-matched male (8-12 weeks) C57BL/6 mice were administered with three once-daily intravenous injections of control (non-specific) or FECH targeting siRNAs (25 μg in 0.5 ml of PBS). In one group (mock) 0.5 ml of PBS was injected. One day later mice were treated with either vehicle or 20 mg/kg vemurafenib (p.o, b.i.d.) for 3 days followed by endpoint analysis of renal function. (A) Representative blots and (B) densitometric analysis showed that the targeted siRNA was able to knock-down FECH proteins levels by approximately 75%. Blots are representative of three independent experiments, all producing similar results. Blood urea nitrogen (C), serum creatinine (D), and histological analysis (E) Renal Ngal gene expression analysis (F) showed that the FECH knockdown mice developed vemurafenib nephrotoxicity within 3 days of treatment, while the control group demonstrated no obvious renal injury or damage. In all the bar graphs (n=6-10 biologically independent samples), experimental values are presented as mean ± s.d. The height of error bar = 1 s.d. and p < 0.05 was indicated as statistically significant. One-way ANOVA followed by Tukey’s multiple-comparison test or non-parametric Mann–Whitney U test was carried out and statistical significance is indicated by *p < 0.05, **p < 0.01, ***p < 0.001.