Extended Data Fig. 3: Global Aldob knockout (KO) promotes tumorigenesis in mice.
a, A 17 bp deletion (from 176 to192) of Aldob gene destroys Aldob protein expression using TALENs technique, and Western Blot of Aldob expression. b, WT and Aldob KO mice fed a fructose free diet do not show weight difference during the experimental period (n = 12 male mice per group). c, Aldolase enzymatic activity in WT and Aldob KO liver tissues (n = 3 biologically independent samples). d, Aldolase enzymatic activity in human HCC tissues (n = 3 biologically independent samples). e, Representative images of WT and Aldob KO livers and H&E histology of tumor nodules at 28th week (scale bars, 50 μm). f, Liver/body weight at 28th week (n = 8 male mice per group). g, Incidence of tumor with >5 mm in WT and Aldob KO mouse livers at 40th week (n = 12 male mice per group). h-i, Representative images (h) and IHC quantification of Aldob, Ki67, VEGFA and β-catenin expressions (i) (n = 10 mice per group, average of 3 IHC slices from each mouse used for statistical analysis) in mouse liver at 40th week (IHC score = intensity x area. The intensity was graded as 0, 1, 2, 3. The area of positive staining was assessed by 0, 1-25%, 26-50%, 51-75%, 76-100%, corresponding to 0, 1, 2, 3, 4 grades, respectively, scale bars, 50 μm). j, Differential metabolite analysis using untargeted metabolomics on WT and Aldob KO mice tumor at the age of 40th week by Triple TOF 6600 MS (n = 6 WT mice, n = 5 KO mice). k, Pathway enrichment analysis in metabolomics (n = 6 WT mice, n = 5 KO mice). l, Relative GSH/GSSG ratios using LC-MS (n = 7 male mice per group). b-d, f, i, l, Data represent the mean ± SD. Statistical significance was determined by two-way ANOVA (b), or two-tailed unpaired t-test (c, d, f, i, l). a, e, Experiments were repeated with three times independently with similar results; data of one representative experiment are shown. Unprocessed immunoblots are shown in Source Data Extended Data Fig. 3.
