Extended Data Fig. 7: Aldob inhibits G6PD activity and enhances p53-mediated G6PD inhibition.
a, Immunofluorescent staining shows co-localization of Aldob and G6PD in Huh7 Aldob cells (scale bars, 10 μm). b, Overview of Aldob and G6PD crystal structures and the potential interaction sites through charge-charge interactions. Dotted lines indicate an outstanding positively charged region on the Aldob surface and a negatively charged region on the G6PD surface. c, The enzymatic activity of purified recombinant Aldob and its mutants (n = 3 biologically independent samples). d, The effect of Aldob and its mutations on G6PD enzyme activity (n = 3 biologically independent samples). e, Western Blot analysis of Aldob expressions in stable Huh7-vector, Huh7-Aldob, Huh7-Aldob-mutant cell lines. f, Protein interaction measurement between G6PD and Aldoa or Aldoc. g, Protein expression and G6PD enzyme activity measurement after Aldoa overexpression (n = 5 biologically independent samples). h, GST pull-down of recombinant GST-p53, His-Aldob and His-G6PD. i, IP assays are performed in exogenous flag-tagged p53 and myc-tagged G6PD transfected into Huh7-vector, Huh7-Aldob and Huh7-Aldob-mutant cells. j, IP assays are performed using G6PD and p53 antibody in WT and p53 KD mice (p53 knockdown mice). c, d, g, Data represent the mean ± SD. Statistical significance was determined by two-tailed unpaired t-test (c, d, g). a, c-k, Experiments were repeated with five times independently with similar results; data of one representative experiment are shown. Unprocessed immunoblots are shown in Source Data Extended Data Fig. 7.
