Fig 2: Serum biomarker levels for AAV-MDR3-Aco-treated Abcb4-/- mice through 12 weeks.The therapeutic effect of AAV treatment is shown in males (a) and females (b & c) that were treated with AAV-MDR3-Aco at 3 × 1013 VG/kg one time (1×, pink, open triangles; n = 3 M (males)/1 F (females)), at 1 × 1014 VG/kg one time (1×, red, filled circles; n = 7 M/6 F), or at 1 × 1014 VG/kg two times (2×, green, filled diamonds; n = 5 F) using as controls saline-treated (blue, open squares; n = 4 M/8 F) and untreated Abcb4+/+ (WT) mice (black, open circles; n = 3 M/3 F). Data are presented as mean + SD. ALP alkaline phosphatase, ALT alanine transaminase, AST aspartate transaminase, BS bile salts. Source data are provided as a Source Data file.

Fig 3: Biliary PC and evidence of cholestasis for Abcb4-/- mice treated with AAV-MDR3-Aco.At the time of sacrifice, PC concentration in bile (a) and liver weight as a percent of body weight (b) were measured for Abcb4-/- mice treated with AAV-MDR3-Aco at 3 × 1013 VG/kg 1× (pink bars or black squares; n = 3 M/1 F), at 1 × 1014 VG/kg 1× (green bars or green triangles; n = 7 M/5 F), or at 1 × 1014 VG/kg 2× (blue bars or blue inverted triangles; n = 3 M/4 F), as well as saline-treated controls (white bars or open circles; n = 4 M/8 F) and untreated WT mice (grey bars or grey circles; n = 3 M/3 F). Liver sections were stained with picrosirius red to indicate fibrosis (d) and the amount of tissue with red staining was quantified (c). Representative pictures from one mouse in each group are shown, except for females treated 1× at 1 × 1014 VG/kg, for which one animal each from both responder (left image) and non-responder (right image) groups is shown. Scale bar = 500 µm. Statistics (one-way ANOVA/Tukey’s multiple comparisons test): *, p < 0.05; **, p < 0.01; ***, p < 0.001; data are presented as mean ± SD; F values and degrees of freedom (numerator, denominator): a males: F(3,12) = 14.52, females: F(3,15) = 4.58; b males: F(3,13) = 8.548, females: F(3,14) = 4.501; c males: F(3,13) = 23.13, females: F(3,14) = 11.58. Source data are provided as a Source Data file.

Fig 4: Methylation of ABCB4 in primary tumors and normal tissues.Representative COBRA analysis is shown for normal human bronchial epithelial cells (NHBEC), human mammary epithelial cells (HMEC passage 2 and 7) and normal human fibroblast (HF-54) and primary non small cell lung cancer (NSCLC), head and neck cancer (HNC) and breast cancer (BrCa). Mock digest (-) and TaqI digest (+) are shown. Products were resolved on 2% gel with 100 bp marker (M). (T = tumor, N = corresponding normal tissue, ivm = in vitro methylated control, m = methylated, u = unmethylated).

Fig 5: Serum biomarker levels for Abcb4-/- mice treated at 5 weeks of age with AAV-MDR3-Aco.The therapeutic effect of AAV treatment is shown in males (a) and females (b) that were treated with AAV-MDR3-Aco at 1 × 1014 VG/kg one time (1×, red, filled circles; n = 4 M/4 F), using as controls saline-treated (blue, open squares; n = 3 M/4 F) and untreated Abcb4+/+ (WT) mice (black, open circles; n = 3 M/3 F). Data are presented as mean + SD. ALP alkaline phosphatase, ALT alanine transaminase, AST aspartate transaminase, BS bile salts. Source data are provided as a Source Data file.