Fig 2: Antiviral activity of ciclopirox in a humanized liver mouse model of HBV infection. a Schematic depiction of the experiment. PXB mice (n = 3 per group) were injected via the tail vein with HBV virions (5 × 107 copies per mouse, genotype D). After 6 weeks, the mice were treated orally with TDF and/or ciclopirox (both 5 mg per kg, daily), and serum samples were taken from the orbital sinus every week. b Serum HBV genomic DNA levels measured by quantitative PCR. c Serum HBsAg levels determined by ELISA. d Serum HBeAg levels determined by ELISA. e Serum ALT levels determined by ELISA. f Human albumin levels determined by ELISA. g Intrahepatic cccDNA was determined in nucleic acid extracted from liver specimens obtained after 35 days of treatment. h Immunohistochemical analysis of hepatitis B core antigen (HBcAg) in liver sections obtained after 35 days of treatment (×200 magnification). The data in b–g are representative of three independent experiments, and are expressed as means ± SDs. The error bars represent the ± SD. Scale bars, 100 μm.*p < 0.05; **p < 0.01, by unpaired two-tailed Student’s t-tests. Source data are provided as a Source Data file

Fig 3: Detection of drug concentration-dependent toxicity and liver injury.Effect of Cloazpine (closed circles) or olanzapine (open circles) on albumin production (a), ALT release (b), and morphology score (c); Effect of troglitazone (closed circles) or pioglitazone (open circles) on albumin production (d), ALT release (e), and morphology score (f); Effect of trovafloxacin (closed circles) or levofloxacin (open circles) on albumin production (g), ALT release (h) and morphology score (i); Immunofluorescence microscopic images showing concentration-dependent increases in caspase 3/7 staining (green;) indicative of apoptosis after treatment with trovafloxacin at 0,1, 10, and 100 (j) times the unbound human Cmax for 7 days; concentration-dependent decrease in TMRM staining (yellow) indicative of mitotoxicity in response to treatment with sitaxsentan at 0,1,10, and 100 (k) times the unbound human Cmax for 7 days. Scale bar represents 50 µm.

Fig 4: MTT assay of nUO and UO hydrolysates on HepG2 cells (A). Assessment of AST and ALT release in HepG2 cells supernatant after 24 h of treatment with nUO and UO (B–C). Bars represent the mean ± s.d. of two independent experiments performed in triplicate and statistically analyzed by One-way Anova followed by Tukey's post-hoc test. C: control, ns: not significant.

Fig 5: Comparison of Drug-Induced AST Release in human hepatocyte Monolayers, liver spheroids, and 3D Liver tissue models. Human hepatocyte monolayers, liver spheroids, and 3D liver tissue models were compared for drug toxicity. The 3D liver tissue models were cultured under air-liquid interface (ALI) conditions for 14 days in tissue culture inserts. Following this, the models were exposed to test drugs every other day over a 7-day period. For all model systems duplicate wells per drug/concentration (N = 2 was used. On days 2, 4, and 7 culture supernatants were collected and released ALT levels were quantified using an ALT specific ELISA kit. The results were then compared across the three model systems to assess differences in drug-induced AST release. There was no difference among the treatment groups (p value < 0.08, Two-way ANOVA) at day 7 post exposure.