Extended Data Fig. 9: Metabolic conditions alter the Pik3caH1047R/wt fitness in vivo.
From: Organismal metabolism regulates the expansion of oncogenic PIK3CA mutant clones in normal esophagus
a-b, Cre-RYFP and Cre-Pik3caH1047R-YFP/wt mice were induced and treated with or without metformin (MET). First suprabasal layer cells per clone (a) and basal clone size distributions (b) analyzed at 28 days. Dots indicate mean and lines S.D. in b. Two-tailed unpaired t-test for suprabasal cells per clone. Two-tailed Kolmogorov-Smirnov test to compare wild-type and mutant basal clone distributions. Contrast ART-C Post-hoc test of differences of differences between distributions. n = 431-917 clones from 5-10 animals per condition. c, Lactate secretion in wild-type cells in minimal medium in control conditions (CTL) or with 2.5 mM MET. Bars indicate mean. Two-tailed paired t-test. d-f, In vitro competition assay. d, Protocol: Confluent cultures of WT-RYFP and Pik3caH1047R/wt cells in minimal medium with/without MET for 28 days. e, Representative confocal image of basal layer of culture from d. WT-RYFP cells, yellow, DAPI, blue. Scale bar, 20 μm. f, Flow cytometric analysis of cultures in d. Each dot represents a biological replicate, lines connect mean values (n = 3–12). Two-tailed paired t-test. g, Urine glucose levels in mice from Fig. 6d. Two-tailed Mann-Whitney test. n = 4 mice per group. Mean ± SD. h, Basal cells per clone in Fig. 6d. Dots and bars indicate mean and S.D. Two-tailed Kolmogorov-Smirnov and Contrast ART-C Post-hoc tests. i, Average proportion of suprabasal cells per clone for each strain and treatment from Fig. 6d, only the first suprabasal cell layer was counted. Each dot corresponds to one animal. Bars are mean ± SD. n = 431-917 clones from 5-10 animals per condition. Two-tailed unpaired t-test. j, Body weight measured at weeks 0 and 4 post diets. Each dot corresponds to one animal, lines link weights from same animal (n = 10-16 mice). Two-tailed paired t-test. k, Blood insulin levels at the end of experiment in j. Error bars are mean±s.e.m. Two-tailed unpaired t-test. l, Average proportion of suprabasal cells per clone for each strain and treatment from Fig. 6g, only first suprabasal cells were counted. Each dot corresponds to one animal. Bars are mean ± S.D. Two-tailed unpaired t-test. m, Distribution of basal cells per clone from Fig. 6g. Dots and bars indicate mean and S.D. Two-tailed Kolmogorov-Smirnov and Contrast ART-C Post-hoc tests.
