Fig 2: ATP7A is a potential oncogene in breast cancer. A, The schematic model of ATP7A/ATP7B regulation of cancer cell cisplatin‐resistance. B, Pearson correlation analysis between mRNA levels of ATP7A and ATP7B in BRCA samples (data came from ENCORI, cBioPortal, LinkedOmics and GEPIA). C, Analysis of ATP7A and ATP7B expression in BRCA and corresponding normal tissues using GEPIA and linkedOmics database. D, Survival curves analysis for breast cancer patients with low versus high expression levels of ATP7A and ATP7B. E, Heat map shows the expression of ATP7A and ATP7B along samples in risk groups. Low expression is represented in green grades and high expression in red grades; box plots shows the expression of ATP7A and ATP7B genes in risk groups, including the P‐value testing for difference using t‐test. F, Enrichment analysis of ATP7A and ATP7B related genes, respectively

Fig 3: ATP7A is an important target of miR‐148a. A, Expression levels of miR‐148a‐3p targets in MDA‐MB‐231 cells transfected with miR‐148a‐3p mimic were determined by RT‐PCR. B, PCR was used to validate mRNA levels altered through overexpression of the corresponding siRNAs. C, Caspase‐3 activity analysis of MDA‐MB‐231 cells treated with cisplatin and multiple siRNAs, respectively, by western blot. D, Caspase‐3 activated probe was used to detect apoptosis in MDA‐MB‐231 cells treated with cisplatin and multiple siRNAs, respectively

Fig 4: MiR‐148a‐3p restrains cisplatin‐resistance through the regulation of ATP7A in breast cancer cells. A, MDA‐MB‐231 cells were co‐transfected with miR‐148a‐3p or miR‐nc and empty or ATP7A‐overexpressed vectors; cell viability was determined using CCK‐8 assays. B, The expression level of miR‐148a‐3p in cisplatin‐resistant cell lines, compared with normal parent cells. C, The expression of miR‐148a‐3p in cisplatin‐treated cells, compared with cells treated with DMSO. D, Expression of miR‐148a‐3p was determined in MDA‐MB‐231 and T47D cells with the increasing doses of cisplatin for 48 hours. Each bar in the figure represents the mean ± SEM of triplicates. * P < .05, ** P < .01

Fig 5: Intein-mediated gene therapy effect on preventing hepatocellular damage and liver disease progression in Atp7b−/− mice(A) Serum alanine and aspartate aminotransferase (ALT and AST) levels in Atp7b+/− healthy control mice and Atp7b−/− mice injected with AAV2/8-TBG-eGFP (GFP) or AAV2/8-HLP-5′ATP7B-N-intein and AAV2/8-HLP-C-intein-3′ATP7B at total doses of 5×1012 (int-ATP7B L) or 2×1013 gc/kg (int-ATP7B H). (B) Representative hematoxylin and eosin (left panels) and Sirius red (right panels) staining. Scale bar: 100 μm. (C) Quantification of Sirius red (SR)-positive areas. Data are expressed as percentage over total field area. (D) Quantification of necroinflammatory grading using Ishak’s scoring system. (E) qPCR analysis of fibrosis and inflammation marker genes. One-way ANOVA plus Tukey’s (A and E) or Kruskal-Wallis (B and D) post-hoc test: ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.005; ∗∗∗∗p < 0.0001. Data are shown as average ± SEM.