Figure 3. Biodistribution of Chol- and GN-MCT1-siRNA in the liver.

Male C57BL/6 wild-type mice (16–18 weeks, n=4) were subcutaneously injected with 10 mg/kg of each siRNA, twice within 15 days, while fed a chow diet. Mice were sacrificed on day 15. (A) Chemical structure of the fully chemically modified siRNA that was used for further in vivo studies: Chol-MCT1-siRNA and GN-MCT1-siRNA. (B, E) Primary hepatocytes, (C, F) stellate cells, and (D, G) Kupffer cells were isolated from each mouse using different gravity centrifugations and gradient solutions after the liver perfusion. Slc16a1/Mct1 mRNA expression levels in each cell-type fraction were measured (mean ± SD, t-test, *: p<0.05, **: p<0.01).

Figure 3—figure supplement 1. Biodistribution of GN-MCT1-siRNA and Chol-MCT1-siRNA.

Male C57BL/6 wild-type mice (16–18 weeks, n=4) were subcutaneously injected with 10 mg/kg of siRNAs twice within 15 days. Livers were perfused through inferior vena cava and multiple liver cells were isolated using different gravity centrifugations and gradient solutions. (A, D) Purity of isolated hepatocytes, (B, E) hepatic stellate cells, (C, F) and Kupffer cells was validated with representative marker genes, Alb, Des, and Clec4f expression, respectively (mean ± SD, one-way ANOVA, *: p<0.05, **: p<0.01, ***: p<0.001, ****: p<0.0001). (G, H) MCT1 protein expression levels in multiple fat tissues (inguinal white adipose tissue [iWAT], gonadal white adipose tissue [gWAT], and brown adipose tissue [BAT]) were monitored by immunohistochemistry. % MCT1 positive areas were quantified. (I, J) MCT1 protein expression levels in multiple tissues (heart, lung, kidney, spleen, and intestine) were monitored by immunohistochemistry. % MCT1 positive areas were quantified.