Fig. 1.

Bulevirtide treatment delays clearance and increases levels of plasma bile salts in Oatp1a1^−/−Ldlr^−/− mice. Using CRISPR/CAS9 the mOatp1a1 locus was mutated in order to generate Oatp1a1^−/−Ldlr^−/− mice. A: sgRNA was targeted to exon 2 and 3 of the mOatp1a1 locus and together with Cas9 injected in the cytoplasm of a zygote derived from Ldlr^+/+ and Ldlr^−/− mice, after which the zygote was implanted into the oviducts of foster mice. Breeding was continued to obtain homozygous Oatp1a1^−/−Ldlr^−/− mice. B: Hepatic OATP1A1 abundance was measured in offspring to confirm knockout (KO) (third lane). The second lane served as a KO control. The gallbladder of male and female Oatp1a1^−/−Ldlr^−/− (blue circle) or Oatp1a1^+/+Ldlr^−/− mice (white circle) treated with a single dose of Bulevirtide was cannulated and [³H]sodium taurocholic acid (TC) was administered intravenously to measure [³H]-TC clearance from (C) plasma and (D) bile and to measure (E) bile flow, which was expressed as μL/minute/100 g body weight (n = 3–5/group). Data are presented as means ± SD. ∗Vehicle versus Bulevirtide. ∗ P < 0.05, according to two-way ANOVA and following Šídák’s multiple-comparisons test (C). Fig. 1A was created with BioRender.com.