Fig. 4

Tumoroids derived from inflammation-induced tumors have increased glucose uptake. a Representative images of colonoids derived from mock Min epithelium and tumoroids derived from mock or ETBF Min tumors 8 weeks post-infection. Scale bar, 100 μm. b Total DNA content in wells of mock and ETBF tumoroids as in a collected at the indicated days post-plating. Mean ± SEM. The data is from three independent experiments each with two biological replicates. *P < 0.05. c Polg gene expression in organoids as in a by qPCR relative to epithelium organoids. Mean ± SEM. The data is from three independent experiments each with two biological replicates. *P < 0.05 compared to epithelium. ^#P < 0.05 compared to mock tumoroid. d qMSP of bisulfite-treated DNA from organoids as in a using methylation-specific primers in promoter CpG islands of indicated genes. Bar indicates mean ± SEM. The data is from three independent experiments each with two biological replicates, asterisk and number sign as in c. e DNA from organoids as in a was used in qPCR assays with primer sets specific for two different regions of mitochondrial DNA or the genomic ApoB. Mean ± SEM. The data is from three independent experiments each with two biological replicates, asterisk and number sign as in c. f Glucose uptake by tumoroids derived as in a. 2-DG6P levels were normalized to protein concentrations of the samples to account for differences in cell number. Mean ± SEM. The data is from three independent experiments each with three biological replicates. *P < 0.05. g At the indicated day post-plating, glucose levels in media from wells with tumoroids were subtracted from glucose levels in media from a well with Matrigel, but without cells, and then normalized to DNA concentration of the tumoroids. Mean ± SEM. The data is from three independent experiments each with two biological replicates. *P < 0.05. h Lactate levels in media from wells with tumoroids at the indicated day post-plating normalized to DNA concentration. Mean ± SEM. The data is from three independent experiments each with two biological replicates. *P < 0.05